Functionalized heterocycles as antiviral agents

ABSTRACT

The present invention discloses compounds of Formula (I), or pharmaceutically acceptable salts, thereof: 
                         
which inhibit the protein(s) encoded by hepatitis B virus (HBV) or interfere with the function of the HBV life cycle of the hepatitis B virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HBV infection. The invention also relates to methods of treating an HBV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/734,424, filed on Sep. 21, 2018. The entire teachings of the above application are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceutical compositions useful as hepatitis virus replication inhibitors. Specifically, the present invention relates to tetracyclic pyridone compounds that are useful in treating viral infections such as hepatitis B virus (HBV). The invention provides novel tetracyclic pyridone compounds as disclosed herein, pharmaceutical compositions containing such compounds, and methods of using these compounds and compositions in the treatment and prevention of HBV infections.

BACKGROUND OF THE INVENTION

Over 240 million people throughout the world are chronically infected with hepatitis B virus (HBV). Out of this patient population, at least 2 million reside in the United States. For those that are chronically infected, many will develop complications of liver disease from cirrhosis or hepatocellular carcinoma (HCC).

HBV is a member of the Hepadnavirus family, and it is able to replicate through the reverse transcription of an RNA intermediate. The 3.2-kb HBV genome exists in a circular, partially doublestranded DNA conformation (rcDNA) that has four overlapping open reading frames (ORF). These encode for the core, polymerase, envelope, and X proteins of the virus. rcDNA must be converted into covalently closed circular DNA (cccDNA) in cells prior to the transcription of viral RNAs. As rcDNA is transcriptionally inert, cccDNA is the only template for HBV transcription, and its existence is required for infection.

The HBV viral envelope contains a mixture of surface antigen proteins (HBsAg). The HBsAg coat contains three proteins that share a common region that includes the smallest of the three proteins (SHBsAg). The other two proteins, Medium HBsAg (MHBsAg) and Large HBsAg (LHBsAg), both contain a segment of SHBsAg with additional polypeptide segments. SHBsAg, MHBsAg, and LHBsAg can also assemble into a non-infectious subviral particle known as the 22-nm particle that contains the same proteins found around infectious viral particles. As the 22-nm particles contain the same antigenic surface proteins that exist around the infectious HBV virion, they can be used as a vaccine to produce neutralizing antibodies.

In chronically infected patients, the non-infectious 22-nm particles are found in much greater abundance than the infectious virions. As a result, the 22-nm particles are thought to be able to protect the infectious virions from the infected host's immune response. Not only can they serve as infectious decoys, but they also suppress normal functioning of immune cells thereby impairing the host's immune response to HBV. Therefore, reducing the level of subviral particles is a feasible therapeutic approach to treating HBV infections. (Refer to WO2015/13990).

In the clinical setting, a diagnostic marker of chronic HBV infection is high serum levels of HBsAg. In recent years, data have suggested that sustained virologic response (SVR) corresponds with HBsAg decline during early treatment, while sustained exposure to HBsAg and other viral antigens might lead to inept immunogenicity. Patients that display higher decreases in serum HBsAg reached a considerably higher SVR following treatment.

Current treatment options for chronically infected HBV patients are limited in number and scope. They include interferon therapy and nucleoside-based inhibitors of HBV DNA polymerase, namely entecavir and tenofovir. The current standard of care is dedicated to reducing the level of viremia and allowance of liver dysfunction, but is associated with negative side-effects and increase persistence of drug-resistant HBV mutants. A significant shortcoming of current therapies is that they are unable to eliminate hepatic resevoirs of cccDNA, prevent transcription of HBsAg from cccDNA, or limit the secretion of HBsAg into serum that will ultimately stifle the immune response. Although compounds have been reported to reduce serum HBsAg levels, they have not been approved as HBV therapies. (Refer to WO2015/113990, WO2015/173164, WO2016/023877, WO2016/071215, WO2016/128335, WO 2017/140821, WO2019097479, WO2019166951, WO2019123285, WO2018198079, WO2018073753, WO2018047109, WO2019110352, WO2019129681, WO2018087345, WO2018083136, WO2018083106, WO2018083081, WO2017216391, WO2018001952, WO2018001944, WO2016107832, WO2016177655, WO2017017042, WO2017017043. WO2017013046, WO2016128335, WO2016071215, WO2015173164, WO2015113990, WO2018219356, WO2018130152, WO2018154466, WO2019069293, WO2017061466, WO2018181883, WO2018161960, WO2017205115, WO2018144605, WO2018085619, WO2018019297, and WO2018022282).

More effective therapies for chronic HBV infections are needed due to this high unmet clinical need. This invention describes the methods to prepare and methods for use of compounds that are believed to suppress the secretion of subviral particles containing HBsAg. Compounds of this type might be used to treat HBV infections and decrease occurrence of liver disease complications such as cirrhosis or HCC.

There is a need in the art for novel therapeutic agents that treat, ameliorate or prevent HBV infection. Administration of these therapeutic agents to an HBV infected patient, either as monotherapy or in combination with other HBV treatments or ancillary treatments, will lead to significantly improved prognosis, diminished progression of the disease, and enhanced seroconversion rates.

SUMMARY OF THE INVENTION

The present invention relates to novel antiviral compounds, pharmaceutical compositions comprising such compounds, as well as methods to treat or prevent viral (particularly HBV) infection in a subject in need of such therapy with said compounds. Compounds of the present invention inhibit the protein(s) encoded by hepatitis B virus (HBV) or interfere with the life cycle of HBV and are also useful as antiviral agents. In addition, the present invention provides processes for the preparation of said compounds.

The present invention provides compounds represented by Formula (I),

and pharmaceutically acceptable salts, N-oxides, esters and prodrugs thereof, wherein:

Z₁ is O, NR₁, or S;

Z₂ is N or CR₂;

Z₃ is N or CR₃;

Z₄ is N or CR₄;

Z₅ is N or CR₅;

preferably, at least one of Z₂ to Z₅ is not nitrogen; more preferably, at least two of Z₂ to Z₅ are not nitrogen;

R₁ is hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;

R₂, R₃, R₄, and R₅ are each independently selected from:

1) hydrogen;

2) halogen;

3) —NO₂;

4) Cyano;

5) Optionally substituted —C₁-C₈ alkyl;

6) Optionally substituted —C₂-C₈ alkenyl;

7) Optionally substituted —C₂-C₈ alkynyl;

8) Optionally substituted —C₃-C₈ cycloalkyl;

9) Optionally substituted 3- to 8-membered heterocycloalkyl;

10) Optionally substituted aryl;

11) Optionally substituted arylalkyl;

12) Optionally substituted heteroaryl;

13) Optionally substituted heteroarylalkyl;

14) —SR₁₁;

15) —S(O)₂R₁₁;

16) —S(O)₂N(R₁₁)(R₁₂);

17) —C(O)R₁₁;

18) —C(O)OR₁₁;

19) —C(O)N(R₁₁)(R₁₂);

20) —C(O)N(R₁₁)S(O)₂(R₁₂);

21) —N(R₁₁)(R₁₂);

22) —N(R₁₃)C(O)N(R₁₁)(R₁₂);

23) —N(R₁₁)C(O)(R₁₂);

24) —N(R₁₁)C(O)₂(R₁₂);

25) —N(R₁₃)S(O)₂N(R₁₁)(R₁₂);

26) —N(R₁₁)S(O)₂(R₁₂);

27) —OR₁₁;

28) —OC(O)R₁₁;

29) —OC(O)OR₁₁; and

30) —OC(O)N(R₁₁)(R₁₂);

wherein R₁₁, R₁₂, and R₁₃ are each independently selected from hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl. Alternatively, R₁₁ and R₁₂ are taken together with the nitrogen atom to which they attached to form an optionally substituted 3-8 membered heterocyclic containing 0, 1, 2, or 3 double bonds.

Alternatively, Z₂ is CR₂, Z₃ is CR₃, and R₂ and R₃ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Alternatively, Z₃ is CR₃, Z₄ is CR₄, and R₃ and R₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Alternatively, Z₄ is CR₄, Z₅ is CR₅, and R₄ and R₅ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Q₁, Q₂, Q₃, and Q₄ are each independently selected from hydrogen, halo, NR₁₁, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈ cycloalkyl; optionally substituted 3- to 8-membered heterocycloalkyl; optionally substituted aryl; and optionally substituted heteroaryl;

Alternatively, Q₁ is taken together with Q₂ or Q₃ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Alternatively, Q₂ is taken together with Q₄ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Alternatively, Q₃ and Q₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds;

Y₁ is hydrogen, halo, or optionally substituted C₁-C₆ alkyl;

Y₂ is O, NR₁₁, N(OR₁₁), or N(NR₁₁);

Y₃ is —C(O)OR₁₁, —C(O)NHSO₂R₁₁, —C(O)NHSO₂NR₁₁R₁₂, 5-tetrazolyl, or 1,2,4-oxadiazol-3-yl-5(4H)-one; and

Y₄ is hydrogen or optionally substituted methyl;

Alternatively, Y₂ and Y₃ are taken together to form an optionally substituted 5-12 membered heterocyclic ring containing 1, 2, or 3 double bonds;

Each preferred group stated above can be taken in combination with one, any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment, the present invention relates to compounds of Formula (I) as described above, and pharmaceutically acceptable salts thereof.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Y₁ is hydrogen, F, Cl, —CH₃ or —CF₃.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Y₂ is O.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Y₃ is —COOH or —C(O)NHSO₂NR₁₁R₁₂, wherein R₁₁ and R₁₂ are as previously defined.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Y₄ is hydrogen or CH₃.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Z₁ is O, S, NH, or NCH₃.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein R₂, R₃, R₄, and R₅ are each independently selected from hydrogen, halogen, —CN, —CH₃, —CF₃, —CHF₂, —C(O)CH₃, —OCH₃, —OCF₃, —OCHF₂, —OH, —OR₁₁, —NH₂, and —NHR₁₂, wherein R₁₁ and R₁₂ are each independently selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein at least one of R₂, R₃, R₄, and R₅ is selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein at least one of R₂, R₃, R₄, and R₅ is selected from one of the following:

wherein each R₂₁ is independently selected from —CH₃, —CHF₂, —CF₃, -isopropyl, -t-butyl, or one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Q₁ is hydrogen, Cl or F; Q₂ is hydrogen, Cl or F, and Q₃ is hydrogen, Cl or F.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Q₄ is -t-butyl or -isopropyl.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Q₁ is hydrogen; Q₂ is hydrogen; Q₃ is hydrogen; and Q₄ is -t-butyl or -isopropyl.

In certain embodiments, the present invention relates to compounds of Formula (I), and pharmaceutically acceptable salts thereof, wherein Q₄ is taken together with Q₁, or Q₂, and the carbon atoms to which they attached to form an optionally substituted ring selected from below:

In another embodiment, the compound of Formula (I) is represented by Formula (II), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, Z₅, Q₁, Q₂, Q₃, and Q₄ are as previously defined. In certain embodiments, Q₃ and Q₄ are taken together with the carbon atom to which they are attached to form a spiro ring. In certain embodiments, Q₁ and Q₃ are both hydrogen, and Q₂ and Q₄ are taken together with the carbon atoms to which they are attached to form a cis-fused ring.

In another embodiment, the compound of Formula (I) is represented by Formula (III-1) or Formula (III-2), or Formula (III-3), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, R₁, Z₂, Z₃, Z₄, Z₅, Q₁, Q₂, Q₃, and Q₄ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by Formula (IV-1), Formula (IV-2), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by Formula (IV-1a), Formula (IV-2a), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by Formula (V-1) or Formula (V-2), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, R₁, Z₂, Z₃, Z₄, Z₅, Q₁, Q₂, Q₃, R₂, R₃, and Q₄ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by Formula (VI), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ are as previously defined, one V is —O—, —C(O)—, —S—, —S(O)₂—, —NR₂₂— or —C(R₂₂)₂—, and the other Vs are independently —O—, —NR₂₂— or —C(R₂₂)₂—; each R₂₂ is independently hydrogen, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₂-C₆ alkenyl, optionally substituted —C₂-C₆ alkynyl, optionally substituted C₁-C₆ alkoxy; optionally substituted —C₃-C₇ cycloalkyl, optionally substituted 3- to 7-membered heterocyclic, optionally substituted aryl or optionally substituted heteroaryl; n is 0, 1, 2 or 3. In certain embodiments, two adjacent Vs are —C(R₂₂)₂—. Alternatively, two adjacent Vs together form —C(R₂₂)═C(R₂₂)—.

In another embodiment, the compound of Formula (I) is represented by Formula (VI-1) or Formula (VI-2), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, Z₅, and V are as previously defined. Alternatively, two adjacent Vs together form —C(R₂₂)═C(R₂₂)—. R₂₂ is as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (VII-1)˜(VII-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, R₁, Z₂, Z₃, Z₄, Z₅, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (VIII-1)˜(VIII-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, R₁, R₂, Z₃, Z₄, Z₅, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (IX-1)˜(IX-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, R₁, Z₂, R₃, Z₄, Z₅, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (X-1)˜(X-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, R₁, R₂, Z₃, Z₄, Z₅, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (XI-1)˜(XI-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, R₁, Z₂, R₃, Z₄, Z₅, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (XII-1)˜(XII-6), or a pharmaceutically acceptable salt thereof:

wherein Y₁, R₂, R₃, Q₄, and V are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (XIII-1)˜(XIII-2), or a pharmaceutically acceptable salt thereof:

wherein R₂, R₃, and Q₄ are as previously defined.

In another embodiment, the compound of Formula (I) is represented by one of Formulae (XIV-1)˜(XIV-4), or a pharmaceutically acceptable salt thereof:

wherein R₂ and R₃ are as previously defined.

In certain embodiments, the present invention relates to compounds of Formulae (VIII-1)˜(VIII-6), (IX-1)˜(IX-6), (X-1)˜(X-6), (XI-1)˜(XI-6), (XII-1)˜(XII-6), (XIII-1)˜(XIII-2), (XIV-1)˜(XIV-4), and pharmaceutically acceptable salts thereof, wherein R₂ and R₃ are each selected from hydrogen, halogen, —CN, —CH₃, —CF₃, —CHF₂, —C(O)CH₃, —OCH₃, —OCF₃, —OCHF₂, —OH, —OR₁₁, —NH₂, and —NHR₁₂, wherein R₁₁ and R₁₂ are each independently selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds of Formulae (VIII-1)˜(VIII-6), (IX-1)˜(IX-6), (X-1)˜(X-6), (XI-1)˜(XI-6), (XII-1)˜(XII-6), (XIII-1)˜(XIII-2), (XIV-1)˜(XIV-4), or pharmaceutically acceptable salts thereof, wherein R₂ and R₃ are each selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

In certain embodiments, the present invention relates to compounds of Formulae (VIII-1)˜(VIII-6), (IX-1)˜(IX-6), (X-1)˜(X-6), (XI-1)˜(XI-6), (XII-1)˜(XII-6), (XIII-1)˜(XIII-2), (XIV-1)˜(XIV-4), or pharmaceutically acceptable salts thereof, wherein R₂ and R₃ are each selected from one of the following:

wherein each R₂₁ is independently selected from —CH₃, —CHF₂, —CF₃, -isopropyl, -t-butyl, or one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.

Representative compounds of the invention include, but are not limited to, compounds of the following formulas and pharmaceutically acceptable salts thereof, where the identity of the variables for each compound are set forth following each formula.

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; 2═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═C—H; Z₄═C—H Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

Z₁═O; Z₂═N; Z₄═C—H Z₁═O; Z₂═C—H; Z₄═N Z₁═S; Z₂═C—H; Z₄═C—H Z₁═S; Z₂═N; Z₄═C—H Z₁═S; Z₂═C—H; Z₄═N Z₁═N—H; Z₂═C—H; Z₄═C—H Z₁═N—H; Z₂═N; Z₄═C—H Z₁═N—H; Z₂═C—H; Z₄═N

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It will be appreciated that the description of the present invention herein should be construed in congruity with the laws and principles of chemical bonding. In some instances, it may be necessary to remove a hydrogen atom in order to accommodate a substituent at any given location.

It will be yet appreciated that the compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic, diastereoisomeric, and optically active forms. It will still be appreciated that certain compounds of the present invention may exist in different tautomeric forms. All tautomers are contemplated to be within the scope of the present invention.

In one embodiment, the compounds described herein are suitable for monotherapy and are effective against natural or native HBV strains and against HBV strains resistant to currently known drugs. In another embodiment, the compounds described herein are suitable for use in combination therapy.

In another embodiment, the compounds of the invention can be used in methods of modulating (e.g., inhibit, disrupt or accelerate) the activity of HBV cccDNA. In yet another embodiment, the compounds of the invention can be used in methods of diminishing or preventing the formation of HBV cccDNA. In another embodiment, the additional therapeutic agent is selected from core inhibitor, which includes GLS4, GLS4JHS, JNJ-379, ABI-H0731, ABI-H2158, AB-423, AB-506, WX-066, and QL-0A6A; immune modulator or immune stimulator therapies, which includes T-cell response activator AIC649 and biological agents belonging to the interferon class, such as interferon alpha 2a or 2b or modified interferons such as pegylated interferon, alpha 2a, alpha 2b, lamda; or STING (stimulator of interferon genes) modulator; or TLR modulators such as TLR-7 agonists, TLR-8 agonists or TLR-9 agonists; or therapeutic vaccines to stimulate an HBV-specific immune response such as virus-like particles composed of HBcAg and HBsAg, immune complexes of HBsAg and HBsAb, or recombinant proteins comprising HBx, HBsAg and HBcAg in the context of a yeast vector; or immunity activator such as SB-9200 of certain cellular viral RNA sensors such as RIG-I, NOD2, and MDA5 protein, or RNA interence (RNAi) or small interfering RNA (siRNA) such as ARC-520, ARC-521, ARB-1467, and ALN-HBV RNAi, or antiviral agents that block viral entry or maturation or target the HBV polymerase such as nucleoside or nucleotide or non-nucleos(t)ide polymerase inhibitors, and agents of distinct or unknown mechanism including agents that disrupt the function of other essential viral protein(s) or host proteins required for HBV replication or persistence such as REP 2139, RG7834, and AB-452. In an embodiment of the combination therapy, the reverse transcriptase inhibitor is at least one of Zidovudine, Didanosine, Zalcitabine, ddA, Stavudine, Lamivudine, Aba-cavir, Emtricitabine, Entecavir, Apricitabine, Atevirapine, ribavirin, acyclovir, famciclovir, valacyclovir, ganciclovir, valganciclovir, Tenofovir, Adefovir, PMPA, cidofovir, Efavirenz, Nevirapine, Delavirdine, or Etravirine.

In another embodiment of the combination therapy, the TLR-7 agonist is selected from the group consisting of SM360320 (9-benzyl-8-hydroxy-2-(2-methoxy-ethoxy)ad-enine), AZD 8848 (methyl [3-({[3-(6-amino-2-butoxy-8-oxo-7,8-dihydro-9H-purin-9-yl)propyl][3-(4-morpholinyl) propyl] amino Imethyl)phenyl]acetate), GS-9620 (4-Amino-2-butoxy-8-[3-(1-pyrrolidinylmethyl)benzyl]-7,8-dihydro-6(5H)-pteridinone), AL-034 (TQ-A3334), and RO6864018.

In another embodiment of the combination therapy, the TLR-8 agonist is GS-9688.

In an embodiment of these combination therapies, the compound and the additional therapeutic agent are co-formulated. In another embodiment, the compound and the additional therapeutic agent are co-administered.

In another embodiment of the combination therapy, administering the compound of the invention allows for administering of the additional therapeutic agent at a lower dose or frequency as compared to the administering of the at least one additional therapeutic agent alone that is required to achieve similar results in prophylactically treating an HBV infection in an individual in need thereof.

In another embodiment of the combination therapy, before administering the therapeutically effective amount of the compound of the invention, the individual is known to be refractory to a compound selected from the group consisting of a HBV polymerase inhibitor, interferon, viral entry inhibitor, viral maturation inhibitor, distinct capsid assembly modulator, antiviral compounds of distinct or unknown mechanism, and combination thereof.

In still another embodiment of the method, administering the compound of the invention reduces viral load in the individual to a greater extent compared to the administering of a compound selected from the group consisting of a HBV polymerase inhibitor, interferon, viral entry inhibitor, viral maturation inhibitor, distinct capsid assembly modulator, antiviral compounds of distinct or unknown mechanism, and combination thereof.

In another embodiment, administering of the compound of the invention causes a lower incidence of viral mutation and/or viral resistance than the administering of a compound selected from the group consisting of a HBV polymerase inhibitor, interferon, viral entry inhibitor, viral maturation inhibitor, distinct capsid assembly modulator, antiviral compounds of distinct or unknown mechanism, and combination thereof.

It should be understood that the compounds encompassed by the present invention are those that are suitably stable for use as pharmaceutical agent.

Definitions

Listed below are definitions of various terms used to describe this invention. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono- or polycyclic carbocyclic ring system comprising at least one aromatic ring, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system that comprises at least one aromatic ring. Polycyclic aryls can comprise fused rings, covalently attached rings or a combination thereof.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclic aromatic radical having one or more ring atom selected from S, O and N; and the remaining ring atoms are carbon, wherein any N or S contained within the ring may be optionally oxidized. Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. A polycyclic heteroaryl can comprise fused rings, covalently attached rings or a combination thereof.

In accordance with the invention, aromatic groups can be substituted or unsubstituted.

The term “alkyl” as used herein, refers to saturated, straight- or branched-chain hydrocarbon radicals. “C₁-C₄ alkyl,” “C₁-C₆ alkyl,” “C₁-C₈ alkyl,” “C₁-C₁₂ alkyl,” “C₂-C₄ alkyl,” or “C₃-C₆ alkyl,” refer to alkyl groups containing from one to four, one to six, one to eight, one to twelve, 2 to 4 and 3 to 6 carbon atoms respectively. Examples of C₁-C₈ alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chain hydrocarbon radicals having at least one carbon-carbon double bond by the removal of a single hydrogen atom. “C₂-C₈ alkenyl,” “C₂-C₁₂ alkenyl,” “C₂-C₄ alkenyl,” “C₃-C₄ alkenyl,” or “C₃-C₆ alkenyl,” refer to alkenyl groups containing from two to eight, two to twelve, two to four, three to four or three to six carbon atoms respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chain hydrocarbon radicals having at least one carbon-carbon double bond by the removal of a single hydrogen atom. “C₂-C₈ alkynyl,” “C₂-C₁₂ alkynyl,” “C₂-C₄ alkynyl,” “C₃-C₄ alkynyl,” or “C₃-C₆ alkynyl,” refer to alkynyl groups containing from two to eight, two to twelve, two to four, three to four or three to six carbon atoms respectively. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

The term “cycloalkyl”, as used herein, refers to a monocyclic or polycyclic saturated carbocyclic ring or a bi- or tri-cyclic group fused, bridged or spiro system, and the carbon atoms may be optionally oxo-substituted or optionally substituted with exocyclic olefinic double bond. Preferred cycloalkyl groups include C₃-C₁₂ cycloalkyl, C₃-C₆ cycloalkyl, C₃-C₈ cycloalkyl and C₄-C₇ cycloalkyl. Examples of C₃-C₁₂ cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, and the like.

The term “cycloalkenyl”, as used herein, refers to monocyclic or polycyclic carbocyclic ring or a bi- or tri-cyclic group fused, bridged or spiro system having at least one carbon-carbon double bond and the carbon atoms may be optionally oxo-substituted or optionally substituted with exocyclic olefinic double bond. Preferred cycloalkenyl groups include C₃-C₁₂ cycloalkenyl, C₃-C₈ cycloalkenyl or C₅-C₇ cycloalkenyl groups. Examples of C₃-C₁₂ cycloalkenyl include, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl, bicyclo[4.2.1]non-3-en-9-yl, and the like.

As used herein, the term “arylalkyl” means a functional group wherein an alkylene chain is attached to an aryl group, e.g., —CH₂CH₂-phenyl. The term “substituted arylalkyl” means an arylalkyl functional group in which the aryl group is substituted. Similarly, the term “heteroarylalkyl” means a functional group wherein an alkylene chain is attached to a heteroaryl group. The term “substituted heteroarylalkyl” means a heteroarylalkyl functional group in which the heteroaryl group is substituted.

As used herein, the term “alkoxy” employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and isomers. Preferred alkoxy are (C₁-C₃) alkoxy.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic and cycloalkenyl moiety described herein can also be an aliphatic group or an alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contains one or more units of unsaturation, e.g., double and/or triple bonds. Examples of aliphatic groups are functional groups, such as alkyl, alkenyl, alkynyl, O, OH, NH, NH₂, C(O), S(O)₂, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH, S(O)₂NH₂, NHC(O)NH₂, NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂, C(O)NHS(O)₂NH or C(O)NHS(O)₂NH₂, and the like, groups comprising one or more functional groups, non-aromatic hydrocarbons (optionally substituted), and groups wherein one or more carbons of a non-aromatic hydrocarbon (optionally substituted) is replaced by a functional group. Carbon atoms of an aliphatic group can be optionally oxo-substituted. An aliphatic group may be straight chained, branched, cyclic, or a combination thereof and preferably contains between about 1 and about 24 carbon atoms, more typically between about 1 and about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, as used herein, aliphatic groups expressly include, for example, alkoxyalkyls, polyalkoxyalkyls, such as polyalkylene glycols, polyamines, and polyimines, for example. Aliphatic groups may be optionally substituted.

The terms “heterocyclic” or “heterocycloalkyl” can be used interchangeably and referred to a non-aromatic ring or a bi- or tri-cyclic group fused, bridged or spiro system, where (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen, (ii) each ring system can be saturated or unsaturated (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, (v) any of the above rings may be fused to an aromatic ring, and (vi) the remaining ring atoms are carbon atoms which may be optionally oxo-substituted or optionally substituted with exocyclic olefinic double bond. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl, and tetrahydrofuryl. Such heterocyclic groups may be further substituted. Heteroaryl or heterocyclic groups can be C-attached or N-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphatic moiety or the like, described herein can also be a divalent or multivalent group when used as a linkage to connect two or more groups or substituents, which can be at the same or different atom(s). One of skill in the art can readily determine the valence of any such group from the context in which it occurs.

The term “substituted” refers to substitution by independent replacement of one, two, or three or more of the hydrogen atoms with substituents including, but not limited to, —F, —Cl, —Br, —I, —OH, C₁-C₁₂-alkyl; C₂-C₁₂-alkenyl, C₂-C₁₂-alkynyl, —C₃-C₁₂-cycloalkyl, protected hydroxy, —NO₂, —N₃, —CN, —NH₂, protected amino, oxo, thioxo, —NH—C₁-C₁₂-alkyl, —NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl, —NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)— heteroaryl, —C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl, —CONH—C₂-C₈-alkynyl, —CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl, —CONH— heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₈-alkenyl, —OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl, —OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —CO₂—C₁-C₁₂ alkyl, —CO₂—C₂-C₈ alkenyl, —CO₂—C₂-C₈ alkynyl, CO₂—C₃-C₁₂-cycloalkyl, —CO₂-aryl, CO₂-heteroaryl, CO₂-heterocyloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl, —OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl, —OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocyclo-alkyl, —NHC(O)H, —NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl, —NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl, —NHC(O)-heterocyclo-alkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl, —NHCO₂— C₂-C₈-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl, —NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂, —NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₈-alkenyl, —NHC(O)NH—C₂-C₈-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl, —NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂, —NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₈-alkenyl, —NHC(S)NH—C₂-C₈-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl, —NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂, —NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₈-alkenyl, —NHC(NH)NH—C₂-C₈-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl, —NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl, —NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl, —NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl, —C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl, —C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl, —C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl, —S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl, —S(O)-heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl, —SO₂NH—C₂-C₈-alkenyl, —SO₂NH— C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl, —SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl, —NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl, —NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl, —NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH, —S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl, —S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, or methylthio-methyl. It is understood that the aryls, heteroaryls, alkyls, cycloalkyls and the like can be further substituted. In certain embodiments, the substituents are independently selected from halo, preferably Cl and F; C₁-C₄-alkyl, preferably methyl and ethyl; C₂-C₄-alkenyl; halo-C₁-C₄-alkyl, such as fluoromethyl, difluoromethyl, and trifluoromethyl; halo-C₂-C₄-alkenyl; C₃-C₆-cycloalkyl, such as cyclopropyl; —CN; —OH; NH₂; C₁-C₄-alkylamino; di(C₁-C₄-alkyl)amino; and NO₂. It is understood that the aryls, heteroaryls, alkyls, and the like can be further substituted. In some cases, each substituent in a substituted moiety is additionally optionally substituted with one or more groups, each group being independently selected from C₁-C₄-alkyl; CF₃, —F, —Cl, —Br, —I, —OH, —NO₂, —CN, and —NH₂.

The term “halo” or halogen” alone or as part of another substituent, as used herein, refers to a fluorine, chlorine, bromine, or iodine atom.

The term “optionally substituted”, as used herein, means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional group(s) individually and independently selected from groups described herein.

The term “hydrogen” includes hydrogen and deuterium. In addition, the recitation of an atom includes other isotopes of that atom so long as the resulting compound is pharmaceutically acceptable.

The term “hydroxy activating group,” as used herein, refers to a labile chemical moiety which is known in the art to activate a hydroxyl group so that it will depart during synthetic procedures such as in a substitution or an elimination reaction. Examples of hydroxyl activating group include, but not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxyl,” as used herein, refers to a hydroxy group activated with a hydroxyl activating group, as defined above, including mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, for example.

The term “hydroxy protecting group,” as used herein, refers to a labile chemical moiety which is known in the art to protect a hydroxyl group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the hydroxy protecting group as described herein may be selectively removed. Hydroxy protecting groups as known in the art are described generally in T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxy-carbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl, benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy group protected with a hydroxy protecting group, as defined above, including benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups, for example.

The term “hydroxy prodrug group,” as used herein, refers to a promoiety group which is known in the art to change the physicochemical, and hence the biological properties of a parent drug in a transient manner by covering or masking the hydroxy group. After said synthetic procedure(s), the hydroxy prodrug group as described herein must be capable of reverting back to hydroxy group in vivo. Hydroxy prodrug groups as known in the art are described generally in Kenneth B. Sloan, Prodrugs, Topical and Ocular Drug Delivery, (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992).

The term “amino protecting group,” as used herein, refers to a labile chemical moiety which is known in the art to protect an amino group against undesired reactions during synthetic procedures. After said synthetic procedure(s) the amino protecting group as described herein may be selectively removed. Amino protecting groups as known in the art are described generally in T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl, benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino group protected with an amino protecting group as defined above.

The term “leaving group” means a functional group or atom which can be displaced by another functional group or atom in a substitution reaction, such as a nucleophilic substitution reaction. By way of example, representative leaving groups include chloro, bromo and iodo groups; sulfonic ester groups, such as mesylate, tosylate, brosylate, nosylate and the like; and acyloxy groups, such as acetoxy, trifluoroacetoxy and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that is relatively inert to proton activity, i.e., not acting as a proton-donor. Examples include, but are not limited to, hydrocarbons, such as hexane and toluene, for example, halogenated hydrocarbons, such as, for example, methylene chloride, ethylene chloride, chloroform, and the like, heterocyclic compounds, such as, for example, tetrahydrofuran and N-methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethyl ether. Such compounds are well known to those skilled in the art, and it will be obvious to those skilled in the art that individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of aprotic solvents may be found in organic chemistry textbooks or in specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The term “protic solvent,” as used herein, refers to a solvent that tends to provide protons, such as an alcohol, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Such solvents are well known to those skilled in the art, and it will be obvious to those skilled in the art that individual solvents or mixtures thereof may be preferred for specific compounds and reaction conditions, depending upon such factors as the solubility of reagents, reactivity of reagents and preferred temperature ranges, for example. Further discussions of protogenic solvents may be found in organic chemistry textbooks or in specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term “stable,” as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintains the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

The synthesized compounds can be separated from a reaction mixture and further purified by a method such as column chromatography, high pressure liquid chromatography, or recrystallization. As can be appreciated by the skilled artisan, further methods of synthesizing the compounds of the Formula herein will be evident to those of ordinary skill in the art. Additionally, the various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing the compounds described herein are known in the art and include, for example, those such as described in R. Larock, Comprehensive Organic Transformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions thereof.

The term “subject,” as used herein, refers to an animal. Preferably, the animal is a mammal. More preferably, the mammal is a human. A subject also refers to, for example, dogs, cats, horses, cows, pigs, guinea pigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and may include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids. The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers may be prepared from their respective optically active precursors by the procedures described above, or by resolving the racemic mixtures. The resolution can be carried out in the presence of a resolving agent, by chromatography or by repeated crystallization or by some combination of these techniques which are known to those skilled in the art. Further details regarding resolutions can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When the compounds described herein contain olefinic double bonds, other unsaturation, or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers or cis- and trans-isomers. Likewise, all tautomeric forms are also intended to be included. Tautomers may be in cyclic or acyclic. The configuration of any carbon-carbon double bond appearing herein is selected for convenience only and is not intended to designate a particular configuration unless the text so states; thus a carbon-carbon double bond or carbon-heteroatom double bond depicted arbitrarily herein as trans may be cis, trans, or a mixture of the two in any proportion.

Certain compounds of the present invention may also exist in different stable conformational forms which may be separable. Torsional asymmetry due to restricted rotation about an asymmetric single bond, for example because of steric hindrance or ring strain, may permit separation of different conformers. The present invention includes each conformational isomer of these compounds and mixtures thereof.

As used herein, the term “pharmaceutically acceptable salt,” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base function with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentane-propionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and aryl sulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers to esters which hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms. Examples of particular esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethylsuccinates.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of a compound of the present invention formulated together with one or more pharmaceutically acceptable carriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier or excipient” means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, preferably by oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intra-arterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectable.

The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or: a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, eye ointments, powders and solutions are also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

For pulmonary delivery, a therapeutic composition of the invention is formulated and administered to the patient in solid or liquid particulate form by direct administration e.g., inhalation into the respiratory system. Solid or liquid particulate forms of the active compound prepared for practicing the present invention include particles of respirable size: that is, particles of a size sufficiently small to pass through the mouth and larynx upon inhalation and into the bronchi and alveoli of the lungs. Delivery of aerosolized therapeutics, particularly aerosolized antibiotics, is known in the art (see, for example U.S. Pat. No. 5,767,068 to Van Devanter et al., U.S. Pat. No. 5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of which are incorporated herein by reference).

Combination and Alternation Therapy

It has been recognized that drug-resistant variants of HIV, HBV and HCV can emerge after prolonged treatment with an antiviral agent. Drug resistance most typically occurs by mutation of a gene that encodes for a protein such as an enzyme used in viral replication, and most typically in the case of HIV, reverse transcriptase, protease, or DNA polymerase, and in the case of HBV, DNA polymerase, or in the case of HCV, RNA polymerase, protease, or helicase. Recently, it has been demonstrated that the efficacy of a drug against HIV infection can be prolonged, augmented, or restored by administering the compound in combination or alternation with a second, and perhaps third, antiviral compound that induces a different mutation from that caused by the principle drug. The compounds can be used for combination are selected from the group consisting of a HBV polymerase inhibitor, interferon, TLR modulators such as TLR-7 agonists or TLR-9 agonists, therapeutic vaccines, immune activator of certain cellular viral RNA sensors, viral entry inhibitor, viral maturation inhibitor, distinct capsid assembly modulator, antiviral compounds of distinct or unknown mechanism, and combination thereof. Alternatively, the pharmacokinetics, biodistribution, or other parameter of the drug can be altered by such combination or alternation therapy. In general, combination therapy is typically preferred over alternation therapy because it induces multiple simultaneous stresses on the virus.

Preferred compounds for combination or alternation therapy for the treatment of HBV include 3TC, FTC, L-FMAU, interferon, adefovir dipivoxil, entecavir, telbivudine (L-dT), valtorcitabine (3′-valinyl L-dC), β-D-dioxolanyl-guanine (DXG), β-D-dioxolanyl-2,6-diaminopurine (DAPD), and β-D-dioxolanyl-6-chloropurine (ACP), famciclovir, penciclovir, lobucavir, ganciclovir, and ribavirin.

Although the invention has been described with respect to various preferred embodiments, it is not intended to be limited thereto, but rather those skilled in the art will recognize that variations and modifications may be made therein which are within the spirit of the invention and the scope of the appended claims.

Antiviral Activity

An inhibitory amount or dose of the compounds of the present invention may range from about 0.01 mg/Kg to about 500 mg/Kg, alternatively from about 1 to about 50 mg/Kg. Inhibitory amounts or doses will also vary depending on route of administration, as well as the possibility of co-usage with other agents.

According to the methods of treatment of the present invention, viral infections, conditions are treated or prevented in a patient such as a human or another animal by administering to the patient a therapeutically effective amount of a compound of the invention, in such amounts and for such time as is necessary to achieve the desired result.

By a “therapeutically effective amount” of a compound of the invention is meant an amount of the compound which confers a therapeutic effect on the treated subject, at a reasonable benefit/risk ratio applicable to any medical treatment. The therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect). An effective amount of the compound described above may range from about 0.1 mg/Kg to about 500 mg/Kg, preferably from about 1 to about 50 mg/Kg. Effective doses will also vary depending on route of administration, as well as the possibility of co-usage with other agents. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or contemporaneously with the specific compound employed; and like factors well known in the medical arts.

The total daily dose of the compounds of this invention administered to a human or other animal in single or in divided doses can be in amounts, for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1 to 25 mg/kg body weight. Single dose compositions may contain such amounts or submultiples thereof to make up the daily dose. In general, treatment regimens according to the present invention comprise administration to a patient in need of such treatment from about 10 mg to about 1000 mg of the compound(s) of this invention per day in single or multiple doses.

The compounds of the present invention described herein can, for example, be administered by injection, intravenously, intra-arterial, subdermally, intraperitoneally, intramuscularly, or subcutaneously; or orally, buccally, nasally, transmucosally, topically, in an ophthalmic preparation, or by inhalation, with a dosage ranging from about 0.1 to about 500 mg/kg of body weight, alternatively dosages between 1 mg and 1000 mg/dose, every 4 to 120 hours, or according to the requirements of the particular drug. The methods herein contemplate administration of an effective amount of compound or compound composition to achieve the desired or stated effect. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 6 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with pharmaceutically excipients or carriers to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w/w). Alternatively, such preparations may contain from about 20% to about 80% active compound.

Lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptoms, the patient's disposition to the disease, condition or symptoms, and the judgment of the treating physician.

Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination of this invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

When the compositions of this invention comprise a combination of a compound of the Formula described herein and one or more additional therapeutic or prophylactic agents, both the compound and the additional agent should be present at dosage levels of between about 1 to 100%, and more preferably between about 5 to 95% of the dosage normally administered in a monotherapy regimen. The additional agents may be administered separately, as part of a multiple dose regimen, from the compounds of this invention. Alternatively, those agents may be part of a single dosage form, mixed together with the compounds of this invention in a single composition.

The “additional therapeutic or prophylactic agents” include but are not limited to, immune therapies (eg. interferon), therapeutic vaccines, antifibrotic agents, anti-inflammatory agents such as corticosteroids or NSAIDs, bronchodilators such as beta-2 adrenergic agonists and xanthines (e.g. theophylline), mucolytic agents, anti-muscarinics, anti-leukotrienes, inhibitors of cell adhesion (e.g. ICAM antagonists), anti-oxidants (eg N-acetylcysteine), cytokine agonists, cytokine antagonists, lung surfactants and/or antimicrobial and anti-viral agents (e.g. ribavirin and amantidine). The compositions according to the invention may also be used in combination with gene replacement therapy.

Abbreviations

Abbreviations which may be used in the descriptions of the scheme and the examples that follow are: Ac for acetyl; AcOH for acetic acid; Boc₂O for di-tert-butyl-dicarbonate; Boc for t-butoxycarbonyl; Bz for benzoyl; Bn for benzyl; t-BuOK for potassium tert-butoxide; Brine for sodium chloride solution in water; CDI for carbonyldiimidazole; DCM or CH₂C₁₂ for dichloromethane; CH₃ for methyl; CH₃CN for acetonitrile; Cs₂CO₃ for cesium carbonate; CuCl for copper (I) chloride; CuI for copper (I) iodide; dba for dibenzylidene acetone; DBU for 1,8-diazabicyclo[5.4.0]-undec-7-ene; DEAD for diethylazodicarboxylate; DIAD for diisopropyl azodicarboxylate; DIPEA or (i-Pr)₂EtN for N,N,-diisopropylethyl amine; DMP or Dess-Martin periodinane for 1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one; DMAP for 4-dimethylamino-pyridine; DME for 1,2-dimethoxyethane; DMF for N,N-dimethylformamide; DMSO for dimethyl sulfoxide; EtOAc for ethyl acetate; EtOH for ethanol; Et₂O for diethyl ether; HATU for O-(7-azabenzotriazol-1-yl)-N,N,N′,N′,-tetramethyluronium Hexafluoro-phosphate; HCl for hydrogen chloride; K₂CO₃ for potassium carbonate; n-BuLi for n-butyl lithium; DDQ for 2,3-dichloro-5,6-dicyano-1,4-benzoquinone; LDA for lithium diisopropylamide; LiTMP for lithium 2,2,6,6-tetramethyl-piperidinate; MeOH for methanol; Mg for magnesium; MOM for methoxymethyl; Ms for mesyl or —SO₂—CH₃; NaHMDS for sodium bis(trimethylsilyl)amide; NaCl for sodium chloride; NaH for sodium hydride; NaHCO₃ for sodium bicarbonate or sodium hydrogen carbonate; Na₂CO₃ sodium carbonate; NaOH for sodium hydroxide; Na₂SO₄ for sodium sulfate; NaHSO₃ for sodium bisulfite or sodium hydrogen sulfite; Na₂S₂O₃ for sodium thiosulfate; NH₂NH₂ for hydrazine; NH₄Cl for ammonium chloride; Ni for nickel; OH for hydroxyl; OsO₄ for osmium tetroxide; OTf for triflate; PPA for polyphophoric acid; PTSA for p-toluenesulfonic acid; PPTS for pyridinium p-toluenesulfonate; TBAF for tetrabutylammonium fluoride; TEA or Et₃N for triethylamine; TES for triethylsilyl; TESCl for triethylsilyl chloride; TESOTf for triethylsilyl trifluoromethanesulfonate; TFA for trifluoroacetic acid; THF for tetrahydrofuran; TMEDA for N,N,N′,N′-tetramethylethylene-diamine; TPP or PPh₃ for triphenyl-phosphine; Tos or Ts for tosyl or —SO₂—C₆H₄CH₃; Ts₂O for tolylsulfonic anhydride or tosyl-anhydride; TsOH for p-tolylsulfonic acid; Pd for palladium; Ph for phenyl; Pd₂(dba)₃ for tris(diben-zylideneacetone) dipalladium (0); Pd(PPh₃)₄ for tetrakis(triphenylphosphine)-palladium (0); PdCl₂(PPh₃)₂ for trans-dichlorobis-(triphenylphosphine)palladium (II); Pt for platinum; Rh for rhodium; rt for room temperature; Ru for ruthenium; TBS for tert-butyl dimethylsilyl; TMS for trimethylsilyl; or TMSCl for trimethylsilyl chloride.

Synthetic Methods

The compounds and processes of the present invention will be better understood in connection with the following synthetic schemes that illustrate the methods by which the compounds of the invention may be prepared. These schemes are of illustrative purpose, and are not meant to limit the scope of the invention. Equivalent, similar, or suitable solvents, reagents or reaction conditions may be substituted for those particular solvents, reagents, or reaction conditions described herein without departing from the general scope of the method of synthesis.

Illustrated in Scheme 1, compounds such as 9 (Q₁, Q₂, Q₃, Q₄, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a carbon-carbon bond forming reaction with reagent 2, typically mediated by a strong base. Amide 2 can be reacted with reagents such as Q₃-M or Q₄-M, where M is defined as a magnesium or lithium containing species, in a carbon-carbon bond forming reaction to produce intermediate 4. Carbonyl intermediate 4 can be reacted in an amination step (denoted as [Amination]) to produce amine 5. This intermediate 5 can be formylated (denoted as [Formylation]) using reagents including, but not limited to: formic acid and ethyl formate, to produce formamide 6. This intermediate can be subjected to a ring closing reaction (denoted as [Ring Closure]) that is mediated by electrophiles including, but not limited to: POCl₃, POBr₃, SOCl₂, or FeCl₃, to produce imine 7. Intermediate 7 can be reacted in an annulation reaction with compound 8 (R defined as optionally substituted alkyl) and following oxidation, the crude product can be purified via SFC to produce final compound 9, with high enantiopurity.

Illustrated in Scheme 2, compounds such as 6 (Q₁, Q₂, Q₃, Q₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a carbon-carbon bond forming reaction with reagent 2, 3, 4, or 5, typically mediated by a strong base to produce carbonyl intermediate 6.

Illustrated in Scheme 3, compounds such as 2 (Q₁, Q₂, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a amide bond forming reaction (denoted as [Amide]) with N, O-dimethylhydroxylamine (or its equivalent) typically mediated by a coupling reagent including, but not limited to: HATU, or Ghosez' reagent, to produce amide 2.

Illustrated in Scheme 4 (R₂, Q₁, Q₂, Q₃, Q₄, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously, X is Cl, Br, I, —B(OH)₂, —BF₃K, —B(pin), —OTf, —OMs, —ONs, or —OTs). Compounds such as 2 can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a carbon-carbon, carbon-oxygen, carbon-nitrogen, or carbon-sulfur bond forming reaction (denoted as [Coupling]) with a reagent such as R₂—H, R₂—B(OR)₂, R₂—BF₃K, R₂—O—R, R₂—NHR, R₂—S—R, R₂—COOH, R₂—SiR₃, R₂—SO₂—NHR (R as defined previously), typically mediated by a base including, but not limited to: K₂CO₃, Cs₂CO₃, KOAc, NaOtBu, NaOH, KOH, Et₃N, DBU, LiHMDS, or NaH, and a metallic reagent (or reagents) that contain(s), but is not limited to containing: Pd, Cu, Zn, Fe, Ir, Ru, Rh, or Ni, to produce 2.

Illustrated in Scheme 5 (R₃, Q₁, Q₂, Q₃, Q₄, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously, X is Cl, Br, I, —B(OH)₂, —BF₃K, —B(pin), —OTf, —OMs, —ONs, or —OTs). Compounds such as 2 can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a carbon-carbon, carbon-oxygen, carbon-nitrogen, or carbon-sulfur bond forming reaction (denoted as [Coupling]) with a reagent including, but not limited to: R₂—H, R₂—B(OR)₂, R₂—BF₃K, R₂—O—R, R₂—NHR, R₂—S—R, R₂—COOH, R₂—SiR₃, R₂—SO₂—NHR (R as defined previously), typically mediated by a base including, but not limited to: K₂CO₃, Cs₂CO₃, KOAc, NaOtBu, NaOH, KOH, Et₃N, DBU, LiHMDS, or NaH, and a metallic reagent (or reagents) that contain(s), but is not limited to containing: Pd, Cu, Zn, Fe, Ir, Ru, Rh, or Ni, to produce 2.

Illustrated in Scheme 6, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₁, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; W₁ defined as optionally substituted alkyl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in saponification reaction with NaOH, and the crude product can be purified via SFC to produce carboxylic acid 9, with high enantiopurity.

Illustrated in Scheme 7, compounds such as 8 (Q₁, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; V₁ and V₂ defined as optionally substituted alkyl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 (W₁ defined as hydrogen or halogen) can be reacted in a carbon-carbon bond forming reaction with nitroalkane 2, typically mediated by a copper source (denoted as [Copper]) including, but not limited to: CuI, CuBr, or Cu₂O, and a ligand (denoted as [Ligand]) including, but not limited to: triethylamine or N,N′-Dimethylcyclohexane-1,2-diamine, to produce diol 3. The nitro function of 3 can be reduced in a redox reaction (denoted as [Reduction]) using reducing mixtures including, but not limited to: Zn/HOAc or Fe/NH₄Cl, to produce amine 4. The amine can be condensed with pyran 5 (Y₁, Y₂, Y₃, and Y₄ as defined previously) to produce diol 6. Diol 6 can be reacted in a ring closing reaction (denoted as [Ring Closure]) that is mediated by electrophilic reagents including, but not limited to: MsCl, PhNTf₂, or Ac₂O. Compound 8 can be produced from intermediate 7 in a carbon-carbon bond forming reaction (denoted as [Coupling]) typically mediated by a base including, but not limited to: K₂CO₃, Cs₂CO₃, KOAc, NaOtBu, NaOH, KOH, Et₃N, DBU, LiHMDS, or NaH, and a metallic reagent (or reagents) that contain(s), but is not limited to containing: Pd, Cu, Zn, Fe, Ir, Ru, Rh, or Ni.

Illustrated in Scheme 8, compounds such as 6 (Q₁, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; V₁ and V₂ defined as optionally substituted alkyl, W₁ defined as hydrogen or optionally substituted carbonyl, W₂ defined as hydrogen or optionally substituted alkoxy) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a carbon-carbon bond forming reaction with nitroalkane 2, typically mediated by a copper source (denoted as [Copper]) including, but not limited to: CuI, CuBr, or Cu₂O, and a ligand (denoted as [Ligand]) including, but not limited to: triethylamine or N,N′-Dimethylcyclohexane-1,2-diamine, to produce diol 3. The nitro function of 3 can be reduced in a redox reaction (denoted as [Reduction]) using reducing mixtures including, but not limited to: Zn/HOAc or Fe/NH₄Cl, to produce amine 4. The amine can be functionalized (denoted as [Protection]) with an electrophilic reagent including, but not limited to: Boc₂O, formic acid, or ethyl formate, to produce intermediate 5. This can undergo a ring closing reaction (denoted as [Ring Closure]) that is mediated by an electrophilic reagent including, but not limited to: MsCl, PhNTf₂, or Ac₂O, to produce heterocycle 6.

Illustrated in Scheme 9, compounds such as 6 (Q₁, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; V₁ and V₂ defined as optionally substituted alkyl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 (W₁ defined as hydrogen or optionally substituted carbonyl, W₂ defined as hydrogen or optionally substituted alkoxy) can be reacted in a halogenation reaction (denoted as [Halogenation]) with an electrophilic reagent including, but not limited to: Br₂, NBS, I₂, or NIS to produce intermediate 2 (W₃ defined as halogen). The amine can undergo a deprotection reaction (denoted as [Deprotection]), under acidic or basic conditions, to produce primary amine 3. The amine can be condensed with pyran 4 (Y₁, Y₂, Y₃, and Y₄ as defined previously) to produce 5. Compound 6 can be produced from intermediate 5 in a carbon-carbon bond forming reaction (denoted as [Coupling]) typically mediated by a base including, but not limited to: K₂CO₃, Cs₂CO₃, KOAc, NaOtBu, NaOH, KOH, Et₃N, DBU, LiHMDS, or NaH, and a metallic reagent (or reagents) that contain(s), but is not limited to containing: Pd, Cu, Zn, Fe, Ir, Ru, Rh, or Ni.

Illustrated in Scheme 10, compounds such as 4 (Q₁, Y₁, Y₂, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; V₁ and V₂ defined as optionally substituted alkyl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be subjected to a ring closing reaction (denoted as [Ring Closure]) that is mediated by electrophiles including, but not limited to: POCl₃, POBr₃, SOCl₂, or FeCl₃, to produce imine 2. This can be reacted in an annulation reaction with compound 3 (R defined as optionally substituted alkyl) and following oxidation with a reagent including, but not limited to: chloranil or DDQ, 4 can be obtained.

Illustrated in Scheme 11, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Imine 1 can be reacted in an annulation reaction with compound 2 (R and V₁ defined as optionally substituted alkyl) and following oxidation with a reagent including, but not limited to: chloranil or DDQ, 3 can be obtained.

Illustrated in Scheme 12, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Pyridone 1 can be reacted in a fluorination reaction (denoted as [Fluorination]) with an electrophilic reagent including, but not limited to: NFSI or SelectFluor, to produce fluorinated compound 2.

Illustrated in Scheme 13, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; X defined as halogen) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acid 1 can be reacted in a halogenation reaction (denoted as [Halogenation]) with an electrophilic reagent including, but not limited to: SOCl₂, oxalyl chloride, Ghosez' Reagent, or POBr₃ to produce acyl halide 2.

Illustrated in Scheme 14, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with hydrazinecarbothioamide and NaOH to produce 2.

Illustrated in Scheme 15, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with hydrazine and trimethylorthoformate to produce 2.

Illustrated in Scheme 16, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, an Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with 2 to produce 3.

Illustrated in Scheme 17, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a functionalization reaction with ammonium hydroxide and palladium (II) chloride to produce 2.

Illustrated in Scheme 18, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with 2, and following treatment with Lawesson's reagent, compounds such as 3 are produced.

Illustrated in Scheme 19, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with ammonium hydroxide and palladium (II) chloride, followed by reaction with sodium azide to produce 2.

Illustrated in Scheme 20, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a cyclization reaction with ammonium hydroxide and DMF-DMA, followed by reaction with hydrazine to produce 2.

Illustrated in Scheme 21, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acyl halide 1 (X defined as halogen) can be reacted in a reaction with sulfone 2 to produce 3.

Illustrated in Scheme 22, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acid 1 can be reacted in a rearrangement reaction with DPPA, a suitable base (denoted as [Base]) including, but not limited to: Et₃N or Hunig's Base, and followed by reaction with HCl, compounds such as 2 are produced.

Illustrated in Scheme 23, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a reaction with acyl halide 2 (X defined as halogen) to produce 3.

Illustrated in Scheme 24, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a reaction with acyl halide 2 (X defined as halogen) to produce 3.

Illustrated in Scheme 25, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a cyclization reaction with dihydrofuran-2,5-dione to produce 3.

Illustrated in Scheme 26, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a reaction with isocyanate 2 to produce 3.

Illustrated in Scheme 27, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a reaction with sulfonyl chloride 2 to produce 3.

Illustrated in Scheme 28, compounds such as 4 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; X₃ defined as halogen) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Amine 1 can be reacted in a Sandmeyer-type reaction with protic compound 2 (X₁ defined as halogen), sodium nitrite, and a potassium salt (X₂ defined as halogen) to produce 3.

Illustrated in Scheme 29, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Halide 1 can be reacted in a borylation reaction, typically mediated by a Pd-containing reagent (denoted as [Pd source]) including, but not limited to: Pd(OAc)₂, PdCl₂(dppf), or Pd(PPh₃)₄, and a base (denoted as [Base]) including, but not limited to: KOAc, K₂CO₃, or Et₃N to produce 2.

Illustrated in Scheme 30, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Boronic acid 1 can be reacted in a coupling reaction with halide 2, typically mediated by a Pd-containing reagent (denoted as [Pd source]) including, but not limited to: Pd(OAc)₂, PdCl₂(dppf), or Pd(PPh₃)₄, and a base (denoted as [Base]) including, but not limited to: KOAc, K₂CO₃, or Et₃N to produce 3.

Illustrated in Scheme 31, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Halide 1 can be reacted in a coupling reaction with 2 (M defined as a functional group containing an atom including, but not limited to: B, Sn, Al, Si, Zn, or Mg), typically mediated by a Pd-containing reagent (denoted as [Pd source]) including, but not limited to: Pd(OAc)₂, PdCl₂(dppf), or Pd(PPh₃)₄, and a base (denoted as [Base]) including, but not limited to: KOAc, K₂CO₃, or Et₃N to produce 3.

Illustrated in Scheme 32, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Halide 1 can be reacted in a coupling reaction with a Cu-containing reagent including, but not limited to: CuBr or CuI, and an alcohol and its conjugate base, namely NaOR/ROH to produce 3.

Illustrated in Scheme 33, compounds such as 3 and/or 4 (Q₁, Q₂, Q₃, Q₄, Y₁, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously; R defined as optionally substituted alkyl, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Ester 1 (W₁ defined as optionally substituted alkyl) can be reacted in a substitution reaction with 2 (M defined as a functional group containing an atom including, but not limited to: B, Sn, Al, Si, Zn, or Mg), typically mediated by a Cu-containing reagent including, but not limited to: CuI or CuBr to produce 3 and/or 4.

Illustrated in Scheme 34, compounds such as 5 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Ester 1 (W₁ defined as optionally substituted alkyl) can be activated by thionyl chloride, then reacted with amine 2 (R defined as hydrogen or optionally substituted alkyl) to produce oxime 3. This can undergo saponification with NaOH to produce acid 4. This intermediate can be reacted in a cyclization reaction (denoted as [Annulation]) to produce 5.

Illustrated in Scheme 35, compounds such as 2 (Q₁, Q₂, Q₃, Q₄, Y₂, Y₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Ester 1 (W₁ defined as optionally substituted alkyl) can be reacted in a cyclization reaction with hydrazine to produce 2.

Illustrated in Scheme 36, compounds such as 3 (Q₁, Q₂, Q₃, Q₄, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Acid 1 can be reacted in an activation reaction (denoted as [Activation]) with a chlorination reagent including, but not limited to: oxalyl chloride, Ghosez' reagent, or thionyl chloride to produce acyl halide 2. This intermediate can undergo a substitution reaction with a reagent such as Q₃-M or Q₄-M (M defined as a functional group containing, but not limited to containing: Mg, Li, Zn, Al or Ni, that is mediated by Cu—X (X defined as halogen) to produce 3.

Illustrated in Scheme 37, compounds such as 9 (Q₁, Q₃, Z₁, Z₂, Z₃, Z₄, and Z₅ as defined previously) can be prepared according to the illustrated synthetic methods herein, or by similar methods known to those skilled in the art. Intermediate 1 can be reacted in a bromination reaction (denoted as [Bromination]) typically mediated by an electrophile including, but not limited to bromine or NBS to produce ester 2. This can undergo an alkylation reaction with intermediate 3 to produce intermediate 4. This can undergo a Heck-type annulation (denoted as [Annulation]) mediated by a metal catalyst including, but not limited to: Pd(PPh₃)₄ or Pd(OAc)₂. Ester 5 can be saponified in aqueous basic media to produce acid 6. This can be activated (denoted as [Activation]) with an electrophile including, but not limited to: oxalyl chloride or thionyl chloride, then reacted in an alkylation reaction (denoted as [Alkylation]) with a nucleophilic source including, but not limited to: t-BuMgCl or iPrMgCl to produce ketone 7. This can be condensed with a chiral or achiral amine (denoted as [Amination]) and following reduction, amine 8 can be produced. This can undergo a deprotection reaction (denoted as [Deprotection]) to produce intermediate 9.

All references cited herein, whether in print, electronic, computer readable storage media or other form, are expressly incorporated by reference in their entirety, including but not limited to, abstracts, articles, journals, publications, texts, treatises, internet web sites, databases, patents, and patent publications.

Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art and such changes and modifications including, without limitation, those relating to the chemical structures, substituents, derivatives, formulations and/or methods of the invention may be made without departing from the spirit of the invention and the scope of the appended claims.

Although the invention has been described with respect to various preferred embodiments, it is not intended to be limited thereto, but rather those skilled in the art will recognize that variations and modifications may be made therein which are within the spirit of the invention and the scope of the appended claims.

EXAMPLES

The compounds and processes of the present invention will be better understood in connection with the following examples, which are intended as an illustration only and not limiting of the scope of the invention. Starting materials were either available from a commercial vendor or produced by methods well known to those skilled in the art.

General Conditions:

Mass spectra were run on LC-MS systems using electrospray ionization. These were Agilent 1290 Infinity II systems with an Agilent 6120 Quadrupole detector. Spectra were obtained using a ZORBAX Eclipse XDB-C18 column (4.6×30 mm, 1.8 micron). Spectra were obtained at 298K using a mobile phase of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). Spectra were obtained with the following solvent gradient: 5% (B) from 0-1.5 min, 5-95% (B) from 1.5-4.5 min, and 95% (B) from 4.5-6 min. The solvent flowrate was 1.2 mL/min. Compounds were detected at 210 nm and 254 nm wavelengths. [M+H]⁺ refers to mono-isotopic molecular weights.

NMR spectra were run on a Bruker 400 MHz spectrometer. Spectra were measured at 298K and referenced using the solvent peak. Chemical shifts for ¹H NMR are reported in parts per million (ppm).

Compounds were purified via reverse-phase high-performance liquid chromatography (RPHPLC) using a Gilson GX-281 automated liquid handling system. Compounds were purified on a Phenomenex Kinetex EVO C18 column (250×21.2 mm, 5 micron), unless otherwise specified. Compounds were purified at 298K using a mobile phase of water (A) and acetonitrile (B) using gradient elution between 0% and 100% (B), unless otherwise specified. The solvent flowrate was 20 mL/min and compounds were detected at 254 nm wavelength.

Alternatively, compounds were purified via normal-phase liquid chromatography (NPLC) using a Teledyne ISCO Combiflash purification system. Compounds were purified on a REDISEP silica gel cartridge. Compounds were purified at 298K and detected at 254 nm wavelength.

Example 1: Synthesis of 6-ethyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: An oven-dried vial was charged with diethyl (2-(methoxy(methyl)amino)-2-oxoethyl)phosphonate (1.535 mL, 7.46 mmol) and THF (18 mL). The reaction mixture was cooled in an ice bath and to this was added sodium hydride (179 mg, 7.46 mmol). After stirring at this temperature for 45 minutes, benzofuran-3(2H)-one (1 g, 7.46 mmol) was added as a solution in THF (9 mL). The reaction mixture was then warmed to rt. After 2 h, the reaction mixture was diluted with water (10 mL) and ethyl acetate (15 mL). The crude product was extracted with ethyl acetate (3×10 mL), then concentrated and purified on silica gel with ethyl acetate:hexanes to provide 2-(benzofuran-3-yl)-N-methoxy-N-methylacetamide (818 mg, 50% yield). ESI MS m/z=219.1 [M+H]⁺.

Step 2: An oven-dried vial was charged with 2-(benzofuran-3-yl)-N-methoxy-N-methylacetamide (818 mg, 3.73 mmol) and THF (9 mL). The reaction mixture was cooled in an ice bath and to this was added 2 M EtMgCl in THF (2.0 mL, 4.0 mmol). After stirring at this temperature for 15 minutes, the reaction was quenched with water (15 mL). The product was extracted with ethyl acetate (3×10 mL), then concentrated and purified on silica gel with ethyl acetate:hexanes to provide 1-(benzofuran-3-yl)butan-2-one (280 mg, 40% yield). ESI MS m/z=189.1 [M+H]⁺.

Step 3: An oven-dried vial was charged with 1-(benzofuran-3-yl)butan-2-one (280 mg, 1.49 mmol) and MeOH (6 mL). Ammonium acetate (344 mg, 4.46 mmol) was added followed by sodium cyanoborohydride (280 mg, 4.46 mmol). The reaction mixture was heated to 60° C. for 1 hour. To the mixture was added 2 M aq. NaOH (3 mL). The product was extracted with dichloromethane (3×15 mL), concentrated, and taken onto the next step.

Step 4: The crude product from the previous step was dissolved in ethyl formate (4 mL, 49.2 mmol). The mixture was heated to 80° C. for 15 min. The mixture was concentrated, and the product was purified on silica gel with 0-10% MeOH:DCM to provide N-(1-(benzofuran-3-yl)butan-2-yl)formamide (227 mg, 70% yield over two steps).

Step 5: An oven-dried vial was charged with N-(1-(benzofuran-3-yl)butan-2-yl)formamide (227 mg, 1.05 mmol) and MeCN (3 mL). POCl₃ (0.1 mL, 1.05 mmol) was added. The reaction mixture was heated to 60° C. for 1 hour. The reaction mixture was allowed to reach room temperature and then quenched with conc. aq. NH₄OH (5 mL). The mixture was concentrated and taken onto the next step.

Step 6: The crude product from the previous step was dissolved in ethanol (2 mL) and water (0.6 mL). Then, ethyl (Z)-2-(ethoxymethylene)-3-oxobutanoate (583 mg, 3.13 mmol) was added. The reaction mixture was heated to 75° C. for 1 hour, concentrated, and taken onto the next step.

Step 7: The crude product from the previous step was dissolved in 1,2-dimethoxyethane (5 mL). Then, p-chloranil (257 mg, 1.044 mmol) was added. The reaction mixture was heated to 70° C. for 2 hours, concentrated, and purified on silica gel 0-10% MeOH:DCM to provide ethyl 6-ethyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (25 mg, 7% yield over three steps).

Step 8: An oven-dried vial was charged with ethyl 6-ethyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (25 mg, 0.07 mmol) and MeOH (1 mL). Then, 6 M aq. NaOH (0.5 mL) was added and stirred for 30 minutes. Then, the pH was adjusted to 3 and the product was extracted with dichloromethane (3×5 mL). The combined organic layers were concentrated, and the residue was purified by RPHPLC to provide 6-ethyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (9 mg, 39% yield). ESI MS m/z=310.1 [M+H]⁺.

Example 2: Synthesis of 2-oxo-6-phenyl-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: An oven-dried flask was charged with 1-Phenyl-2-(triphenylphosphoranylidene)ethanone (8.51 g, 22.37 mmol), benzofuran-3(2H)-one (2 g, 14.91 mmol) and toluene (50 mL). The reaction mixture was heated to 120° C. for 18 h, then allowed to reach rt. The reaction mixture was partitioned with water, and extracted with ethyl acetate (3×50 mL), then concentrated and purified on silica gel with ethyl acetate:hexanes to provide 2-(benzofuran-3-yl)-1-phenylethan-1-one (892 mg, 25% yield). ESI MS m/z=237.1 [M+H]⁺.

2-(benzofuran-3-yl)-1-phenylethan-1-one was used to synthesize Example 2 in a manner analogous to the methods used for Example 1. ESI MS m/z=358.1 [M+H]⁺.

Example 3: Synthesis of (S)-6-(tert-butyl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: An oven-dried vial was charged with 2-(benzofuran-3-yl)acetic acid (916 mg, 5.2 mmol) and anhydrous DCM (5 mL). Three drops of anhydrous DMF were added, then the mixture was cooled in an ice bath. Then, oxalyl chloride (726 mg, 5.72 mmol) was added dropwise. The reaction mixture was warmed to rt, concentrated, and used directly in the next step.

Step 2: The material from the previous step was dissolved in THF (10 mL) and put under a nitrogen atmosphere. To this was added CuBr (746 mg, 5.2 mmol) and then the reaction mixture was cooled in an ice bath. Then, a 1.7 M solution of t-butyl magnesium chloride in THF (3.3 mL, 5.7 mmol) was added dropwise. The reaction mixture was allowed to reach rt and stirred overnight. Then, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3×20 mL). The combined organics were concentrated and purified on silica gel with ethyl acetate:hexanes to provide 1-(benzofuran-3-yl)-3,3-dimethylbutan-2-one (650 mg, 58% yield over two steps). ESI MS m/z=217.1 [M+H]⁺.

1-(benzofuran-3-yl)-3,3-dimethylbutan-2-one was used to synthesize Example 3 in a manner analogous to the methods used for Example 1. ESI MS m/z=338.1 [M+H]⁺.

Racemic 6-(tert-butyl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (50 mg) was subjected to purification by SFC (Phenomenex i-Amylose-1 column, 20% MeOH:CO₂ isocratic gradient) to provide (S)-6-(tert-butyl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (10 mg, >99% ee). ESI MS m/z=338.1 [M+H]⁺.

Example 4: Synthesis of (S)-6-(tert-butyl)-11-(difluoromethoxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: An oven-dried flask was charged with 7-hydroxybenzofuran-3(2H)-one (6 g, 40 mmol), DCM (25 mL), (bromodifluoromethyl)trimethylsilane (16.2 g, 80 mmol), NaOH (8 g, 200 mmol), and water (32 mL). The reaction mixture was stirred vigorously for 18 h then organic layer was removed. The product was extracted with DCM (3×25 mL), the combined organic layers were concentrated. The residue was purified on silica gel with ethyl acetate:hexanes to provide 7-(difluoromethoxy)benzofuran-3(2H)-one (1.2 g, 15% yield). ESI MS m/z=201.1 [M+H]⁺.

Step 2: An oven-dried vial was charged with diethyl (2-(methoxy(methyl)amino)-2-oxoethyl)phosphonate (60 mg, 0.25 mmol) and THF (5 mL). The reaction mixture was cooled in an ice bath and to this was added sodium hydride (7 mg, 7.46 mmol). After stirring at this temperature for 45 minutes, 7-(difluoromethoxy)benzofuran-3(2H)-one (50 mg, 0.25 mmol) was added as a solution in THF (3 mL). The reaction mixture was then warmed to rt. After 2 h, the reaction mixture was diluted with water (10 mL) and ethyl acetate (15 mL). The crude product was extracted with ethyl acetate (3×10 mL), then concentrated and purified on silica gel with ethyl acetate:hexanes to provide 2-(7-(difluoromethoxy)benzofuran-3-yl)-N-methoxy-N-methylacetamide (36 mg, 50% yield). ESI MS m/z=286.1 [M+H]⁺.

Step 3: An oven-dried vial was charged with 2-(7-(difluoromethoxy)benzofuran-3-yl)-N-methoxy-N-methylacetamide (36 mg, 0.13 mmol) and THF (5 mL). The reaction mixture was cooled in an ice bath and to this was added 1.7 M solution of t-butyl magnesium chloride in THF (0.1 mL, 0.17 mmol). After stirring at this temperature for 15 minutes, the reaction was quenched with water (15 mL). The product was extracted with ethyl acetate (3×10 mL), then concentrated and purified on silica gel with ethyl acetate:hexanes to provide 1-(7-(difluoromethoxy)benzofuran-3-yl)-3,3-dimethylbutan-2-one (30 mg, 83% yield). ESI MS m/z=283.1 [M+H]⁺.

1-(7-(difluoromethoxy)benzofuran-3-yl)-3,3-dimethylbutan-2-one was used to synthesize Example 4 in a manner analogous to the methods used for Example 1. ESI MS m/z=404.1 [M+H]⁺.

Racemic 6-(tert-butyl)-11-(difluoromethoxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (50 mg) was subjected to purification by SFC (Phenomenex i-Amylose-1 column, 20% MeOH:CO₂ isocratic gradient) to provide (S)-6-(tert-butyl)-11-(difluoromethoxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (9 mg, >99% ee). 1H-NMR (CDCl₃): δ 15.4 (s, 1H), 8.53 (s, 1H), 7.47 (dd, J=7.6, 1.3 Hz), 7.29-7.37 (m, 2H), 7.12 (s, 1H), 6.78 (t, J=72.8 Hz, 1H), 4.21 (d, J=5.9 Hz, 1H), 3.40-3.51 (m, 2H), 0.93 (s, 9H). ESI MS m/z=404.1 [M+H]⁺.

Example 5: Synthesis of (S)-6-(tert-butyl)-10-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: Into a 5-L 4-necked round-bottom flask, was placed (6-hydroxy-1-benzofuran-3-yl)acetic acid (200.00 g, 1 equiv.), MeOH (2000.00 mL), H₂SO₄ (12.00 mL). The resulting solution was stirred for 1 h at 60° C. The resulting mixture was concentrated under vacuum 141. The resulting solution was diluted with 2 L of H₂O. The pH value of the solution was adjusted to 9 with NaHCO₃(sat.). The resulting solution was extracted with 2×2 L of ethyl acetate. The solvent was concentrated under vacuum. This resulted in 200 g (93.20%) of methyl 2-(6-hydroxy-1-benzofuran-3-yl)acetate as an off-white solid.

Step 2: Into a 5-L 4-necked round-bottom flask, was placed methyl 2-(6-hydroxy-1-benzofuran-3-yl)acetate (200.00 g, 969.946 mmol, 1.00 equiv.), DMF (2000.00 mL), K₂CO₃ (201.08 g, 1454.919 mmol, 1.5 equiv.), and methyl iodide (206.51 g, 1454.919 mmol, 1.5 equiv.). The resulting solution was stirred for 16 h at 70° C. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 2 L of H₂O. The resulting solution was extracted with 2×2 L of ethyl acetate. The solvent was concentrated under vacuum. This resulted in 180 g (84.27%) of methyl 2-(6-methoxy-1-benzofuran-3-yl)acetate as a yellow solid.

Step 3: Into a 3-L 4-necked round-bottom flask, was placed methyl 2-(6-methoxy-1-benzofuran-3-yl)acetate (180.00 g, 817.350 mmol, 1.00 equiv.), THF (900.00 mL). This was followed by the addition of a solution of NaOH (65.38 g, 1634.699 mmol, 2.00 equiv.) in H₂O (900 mL) in portions at 0° C. The resulting solution was stirred for 1 h at room temperature. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 2 L of H₂O. HCl (2 mol/L) was employed to adjust the pH to 4. The solids were collected by filtration. This resulted in 160 g (94.94%) of (6-methoxy-1-benzofuran-3-yl)acetic acid as a yellow solid.

Step 4: Into a 5-L 4-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed (6-methoxy-1-benzofuran-3-yl)acetic acid (175.00 g, 848.703 mmol, 1.00 equiv.), DCM (3500.00 mL), and DMF (2.00 mL, 25.844 mmol, 0.03 equiv.). Then (COCl)₂ (118.49 g, 933.573 mmol, 1.1 equiv.) was added dropwise. The resulting solution was stirred for 4 h at room temperature. The resulting mixture was concentrated under vacuum. This resulted in 170 g (89.17%) of (6-methoxy-1-benzofuran-3-yl)acetyl chloride (crude) as green oil.

Step 5: Into a 5-L 4-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed (6-methoxy-1-benzofuran-3-yl)acetyl chloride (170.00 g, 756.766 mmol, 1.00 equiv.), CuBr (108.56 g, 756.766 mmol, 1.00 equiv.), a solution of tert-butyl(chloro)magnesium in THF (1M, 832 mL) was added dropwise. The resulting solution was stirred for 3 h at room temperature. The reaction was then quenched by the addition of 3 L of NH₄Cl (aq.). The resulting solution was extracted with 3×3 L of ethyl acetate. The solution was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (5%). This resulted in 80 g (42.92%) of 1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-one as a light yellow solid.

Step 6: Into a 2-L 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed (S)-phenylethylamine (118.08 g, 974.398 mmol, 3 equiv.), DCE (800.00 mL), TEA (164.33 g, 1623.996 mmol, 5 equiv.), TiCl₄ (46.21 g, 243.599 mmol, 0.75 equiv.) was added dropwise, 1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-one (80.00 g, 324.799 mmol, 1.00 equiv.) was then added. The resulting solution was stirred for 16 h at 80° C. The solids were filtered out. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 1 L of MeOH. NaBH₄ (12.29 g, 324.799 mmol, 1.00 equiv.) was added slowly at 0° C. The resulting solution was stirred for 1 h at 10° C. The reaction was then quenched by the addition of 1 L of NH₄Cl (sat.). The aqueous layer was extracted with EA (2×1 L). The resulting mixture was concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2%). This resulted in 91 g (79.71%) of [(2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl][(1S)-1-phenylethyl]amine as yellow oil.

Step 7: Into a 2-L round-bottom flask was placed [(2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl][(1S)-1-phenylethyl]amine (91.00 g, 1 equiv.), EtOH (900.00 mL), Pd(OH)₂/C (18.00 g). The resulting solution was stirred for 3 h at 30° C. under H₂ atmosphere (2 atm). The solids were filtered out. The residue was applied onto a silica gel column with dichloromethane/methanol (5%). This resulted in 58 g (90.58%) of (2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-amine as light yellow oil.

Step 8: Into a 1-L round-bottom flask, was placed (2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-amine (58.00 g, 234.497 mmol, 1.00 equiv.), ethyl formate (500.00 mL). The resulting solution was stirred for 16 h at 65° C. The resulting mixture was concentrated under vacuum. This resulted in 58 g (89.83%) of N-[(2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl]formamide as yellow oil.

Step 9: Into a 1-L round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed N-[(2S)-1-(6-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl]formamide (58.00 g, 1 equiv.), acetonitrile (600.00 mL), methanesulfonic acid (120.00 mL).

The resulting solution was stirred for 2 h at 80° C. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 1 L of H₂O. The pH value of the solution was adjusted to 10 with Na₂CO₃ (sat.). The resulting solution was extracted with 2×1 L of dichloromethane. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (20%). This resulted in 40 g (73.79%) of (S)-3-(tert-butyl)-7-methoxy-3,4-dihydrobenzofuro[2,3-c]pyridine as a white solid.

Step 10: Into a 1-L round-bottom flask, was placed (S)-3-(tert-butyl)-7-methoxy-3,4-dihydrobenzofuro[2,3-c]pyridine (41.00 g, 159.327 mmol, 1.00 equiv.), EtOH (400.00 mL), TFA (19.98 g, 175.259 mmol, 1.10 equiv.), ethyl (2Z)-2-(ethoxymethylidene)-3-oxobutanoate (89.00 g, 477.980 mmol, 3.00 equiv.). The resulting solution was stirred for 1 h at 80° C. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 500 mL of H₂O. The pH value of the solution was adjusted to 10 with Na₂CO₃ (sat.). The resulting solution was extracted with 3×500 mL of ethyl acetate The residue was applied onto a silica gel column with PE/EA/EtOH (2:1:1). This resulted in 35 g (55.27%) of ethyl (6S)-6-(tert-butyl)-10-methoxy-2-oxo-1,6,7,12b-tetrahydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as an orange syrup.

Step 11: Into a 1-L round-bottom flask, was placed ethyl (6S)-6-(tert-butyl)-10-methoxy-2-oxo-1,6,7,12b-tetrahydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (35.00 g, 88.057 mmol, 1.00 equiv.), DME (350.00 mL), chloranil (25.98 g, 105.668 mmol, 1.20 equiv.). The resulting solution was stirred for 1 h at 80° C. The reaction was then quenched by the addition of 1500 mL of Na₂S₂O₃ (10%). The pH value of the solution was adjusted to 12 with Na₂CO₃(sat.). The resulting solution was extracted with 3×1 L of dichloromethane and the organic layers combined. The residue was applied onto a silica gel column with dichloromethane/EtOH (5%). This resulted in 22 g (63.18%) of ethyl (S)-6-(tert-butyl)-10-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as an orange syrup.

Step 12: Into a 1-L round-bottom flask, was placed ethyl (S)-6-(tert-butyl)-10-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (22.00 g), HBr (40%, aq, 600.00 mL). The resulting solution was stirred for 16 h at 100° C. The resulting mixture was concentrated under vacuum. This resulted in 25 g of (S)-6-(tert-butyl)-10-hydroxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a yellow solid (crude).

Step 13: Into a 1-L round-bottom flask, was placed (S)-6-(tert-butyl)-10-hydroxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (25.00 g, crude), EtOH (300.00 mL), SOCl₂(30.00 mL). The resulting solution was stirred for 16 h at 80° C. The resulting mixture was concentrated under vacuum. The resulting solution was diluted with 2 L of NaHCO₃(10%). The resulting solution was extracted with 4×1 L of dichloromethane. The residue was applied onto a silica gel column with DCM/EtOH (6%). This resulted in 10.2186 g (48.1%) of ethyl (S)-6-(tert-butyl)-10-hydroxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as an off-white solid. 1H-NMR (DMSO-d6): δ 10.03 (s, 1H), 8.44 (s, 1H), 7.62 (d, J=8.5 Hz, 1H), 7.00 (d, J=2.0 Hz, 1H), 6.85 (dd, J=8.5, 2.1 Hz, 1H), 6.36 (s, 1H), 4.51 (d, J=6.8 Hz, 1H), 4.21 (q, J=7.1 Hz, 2H), 3.47-3.37 (m, 1H), 3.28 (d, J=7.0 Hz, 1H), 1.27 (t, J=7.1 Hz, 3H), 0.81 (s, 9H). ESI MS m/z=382.1 [M+H]⁺.

Step 14: A vial was charged with ethyl (S)-6-(tert-butyl)-10-hydroxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (100 mg) and anhydrous DCM (20 mL) under a nitrogen atmosphere. Then, N-Phenyl-bis(trifluoromethanesulfonimide) (500 mg) was added and the reaction was stirred for 8 h at rt. The solvent was removed and the residue was purified on silica gel to provide ethyl (S)-6-(tert-butyl)-2-oxo-10-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (65 mg). ESI MS m/z=514.1 [M+H]⁺.

Step 15: An oven-dried vial was charged with ethyl (S)-6-(tert-butyl)-2-oxo-10-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (50 mg), 2-(furan-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (124 mg), Pd^(t)BuXPhos G3 (10 mg), and Cs₂CO₃ (194 mg). The vial was purged with nitrogen gas for 5 minutes, then DMF (3 mL) and water (0.8 mL) were added via syringe. The reaction mixture was heated under a nitrogen atmosphere for 80 minutes at 110° C. After cooling to room temperature, the reaction mixture was filtered and the product was purified by RPHPLC to provide ethyl (S)-6-(tert-butyl)-10-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as a white solid (15 mg). ESI MS m/z=432.1 [M+H]⁺.

Step 16: A vial was charged with ethyl (S)-6-(tert-butyl)-10-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (15 mg), MeOH (1 mL), THF (1 mL), 1 M aq. NaOH (0.5 mL). The reaction mixture was stirred for 4 h, then the organic solvents were removed under a stream of nitrogen. The pH of the mixture was adjusted to 3 by addition of 1 M aq. HCl. The product was extracted with DCM, then the DCM was removed under a stream of nitrogen. The residue was purified by RPHPLC to provide (S)-6-(tert-butyl)-10-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a white solid (2 mg). ESI MS m/z=404.1 [M+H]⁺.

Example 6: (S)-6-(tert-butyl)-10-(difluoromethoxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 4. ESI MS m/z=404.1 [M+H]⁺.

Example 7: (S)-6-(tert-butyl)-10-(difluoromethoxy)-3-(1H-1,2,4-triazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizin-2-one

Step 1: A vial was charged with (S)-6-(tert-butyl)-10-(difluoromethoxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid (100 mg) and DCM (5 mL). Then it was cooled in an ice bath before PCl₅ (62 mg) was added. After 30 min, ammonium hydroxide (0.12 mL) was added via syringe and the mixture was stirred overnight. The reaction mixture was partitioned with water and the product was extracted with DCM and concentrated. The residue was dissolved in 1,1-dimethoxy-N,N-dimethylmethanamine (1.6 mL) and heated to 95° C. for 45 minutes. After concentrating, the residue was dissolved in HOAc (2.7 mL). Then, hydrazine hydrate (0.06 mL) was added and the reaction mixture was heated at 95° C. for 45 minutes. The volatiles were removed and the product was purified by RPHPLC to provide (S)-6-(tert-butyl)-10-(difluoromethoxy)-3-(1H-1,2,4-triazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizin-2-one as a white solid (15 mg). ESI MS m/z=427.1 [M+H]⁺.

Example 8: (S)-6-(tert-butyl)-11-(difluoromethoxy)-3-(1H-1,2,4-triazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizin-2-one

The title compound was prepared in a manner analogous to the methods used for Example 7. ESI MS m/z=427.1 [M+H]⁺.

Example 9: (S)-6-(tert-butyl)-10-(1-methyl-1H-pyrazol-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=418.1 [M+H]⁺.

Example 10: (S)-6-(tert-butyl)-10-(1-methyl-1H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=418.1 [M+H]⁺.

Example 11: Synthesis of (S)-6-(tert-butyl)-10-(1-methyl-1H-pyrazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=418.1 [M+H]⁺.

Example 12: Synthesis of (S)-6-(tert-butyl)-10-cyano-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=363.1 [M+H]⁺.

Example 13: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(1H-pyrazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=404.1 [M+H]⁺.

Example 14: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(1H-pyrazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=404.1 [M+H]⁺.

Example 15: Synthesis of (S)-6-(tert-butyl)-10-(furan-2-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=404.1 [M+H]⁺.

Example 16: Synthesis of (S)-6-(tert-butyl)-10-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=404.1 [M+H]⁺.

Example 17: Synthesis of (S)-6-(tert-butyl)-10-(oxazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=405.1 [M+H]⁺.

Example 18: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(thiophen-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=420.1 [M+H]⁺.

Example 19: Synthesis of (S)-6-(tert-butyl)-10-(isothiazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=421.1 [M+H]⁺.

Example 20: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(thiophen-3-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=420.1 [M+H]⁺.

Example 21: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(thiazol-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=421.1 [M+H]⁺.

Example 22: Synthesis of (S)-6-(tert-butyl)-2-oxo-10-(thiazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=421.1 [M+H]⁺.

Example 23: Synthesis of (S)-6-(tert-butyl)-10-cyclopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=378.1 [M+H]⁺.

Example 24: Synthesis of (S)-6-(tert-butyl)-10-methyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=352.1 [M+H]⁺.

Example 25: Synthesis of (S)-6-(tert-butyl)-10-ethyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 5. ESI MS m/z=366.1 [M+H]⁺.

Example 26: Synthesis of (S)-6-(tert-butyl)-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: To a stirred solution of ethyl 2-(7-methoxy-1-benzofuran-3-yl)acetate (450.00 g, 1921.016 mmol, 1.00 equiv.) in THF (2.25 L) was added a solution of NaOH (230.51 g, 5763.049 mmol, 3.00 equiv.) dissolved in H₂O (2.25 L). The resulting mixture was stirred for 1 h at room temperature. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 2-3 with 6N HCl. The precipitated solids were collected by filtration and washed with water (500 mL) to afford (7-methoxy-1-benzofuran-3-yl)acetic acid (400 g, crude) as a light yellow solid.

Step 2: To a stirred mixture of (7-methoxy-1-benzofuran-3-yl)acetic acid (400.00 g, 1939.892 mmol, 1.00 equiv.) in DCM (4.00 L) was added (CO)₂Cl₂ (738.67 g, 5819.677 mmol, 3.00 equiv.) dropwise at room temperature. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in THF (4.00 L). To the above mixture was added CuBr (278.28 g, 1939.892 mmol, 1.00 equiv.) and tert-butylmagnesium chloride (226.72 g, 1939.892 mmol, 1.00 equiv.) dropwise over 30 min at room temperature. The reaction was quenched with NH₄C₁ (sat.). The resulting mixture was extracted with EtOAc (2×2 L). The combined organic layers were washed with brine (1×1 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (5:1) to afford 1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-one (230 g, 48.14%) as a dark yellow solid.

Step 3: To a stirred mixture of D-methylbenzylamine (282.90 g, 2334.494 mmol, 2.50 equiv.) and TEA (472.45 g, 4668.989 mmol, 5.00 equiv.) in DCE (2.50 L) was added TiCl₄ (177.12 g, 933.798 mmol, 1.00 equiv.) dropwise at 0° C. under nitrogen atmosphere. After 10 min, to the above mixture was added 1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-one (230.00 g, 933.798 mmol, 1.00 equiv.) dissolved in DCE (500 mL). The resulting mixture was stirred overnight at 70° C. The mixture was allowed to cool down to 0° C. To the above mixture was added NaBH₄ (70.66 g, 1867.596 mmol, 2.00 equiv.) and MeOH (500.00 mL) in portions over. The resulting mixture was stirred for 3 h at room temperature. The reaction was quenched by the addition of Water (500 mL). The mixture was acidified to pH 1-4 with 2N HCl. The resulting mixture was extracted with CH₂Cl₂ (3×1000 mL). The combined organic layers were washed with Na₂CO₃ (aq) (1×2 L) and brine (1×2 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH₂Cl₂/MeOH (20:1) to afford [(2R)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl][(1R)-1-phenylethyl]amine (230 g, 70.07%) as a brown oil.

Step 4: To a solution of [(2R)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl][(1R)-1-phenylethyl]amine (230.00 g, 1 equiv.) in MeOH (2.30 L) was added Pd/C (20.00 g) under nitrogen atmosphere in a 10 L pressure tank reactor. The mixture was hydrogenated at 40° C. for overnight under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH₂Cl₂/MeOH (40:1) to afford (2S)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-amine (150 g, 92.68%) as a colorless oil.

Step 5: Into a 3 L 4-necked round-bottom flask were added (2S)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-amine (150.00 g, 606.458 mmol, 1.00 equiv.) and ethyl formate (1500.00 mL). The resulting mixture was stirred overnight at 70° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure to afford N-[(2R)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl]formamide (160 g, 95.82%) as a dark yellow solid.

Step 6: Into a 3 L 4-necked round-bottom flask were added N-[(2R)-1-(7-methoxy-1-benzofuran-3-yl)-3,3-dimethylbutan-2-yl]formamide (160.00 g, 1 equiv.), methanesulfonic acid (250.00 mL) and ACN (1250.00 mL). The resulting mixture was stirred for 5 h at 80° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure to afford (4S)-4-tert-butyl-10-methoxy-8-oxa-5-azatricyclo[7.4.0.0{circumflex over ( )}[2,7]]trideca-1(9),2(7),5,10,12-pentaene (130 g, 86.94%) as a dark yellow solid.

Step 7: Into a 2 L 4-necked round-bottom flask were added (4S)-4-tert-butyl-10-methoxy-8-oxa-5-azatricyclo[7.4.0.0{circumflex over ( )}[2,7]]trideca-1(9),2(7),5,10,12-pentaene (130.00 g, 505.182 mmol, 1.00 equiv.), TFA (57.60 g, 505.182 mmol, 1.00 equiv.), ethyl (2Z)-2-(ethoxymethylidene)-3-oxobutanoate (282.21 g, 1515.571 mmol, 3.00 equiv.) and EtOH (1.30 L). The resulting mixture was stirred for 2 h at 70° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (1:1) to afford ethyl (8R)-8-tert-butyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),5,11(16),12,14-pentaene-5-carboxylate (80 g, 39.84%) as a brown oil.

Step 8: To a stirred solution of ethyl (8R)-8-tert-butyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10), 5,11(16),12,14-pentaene-5-carboxylate (80.00 g, 201.273 mmol, 1.00 equiv.) in DCM (800.00 mL) was added DDQ (54.83 g, 241.538 mmol, 1.20 equiv.). The resulting mixture was stirred for 1 h at room temperature. The reaction was quenched by the addition of Na₂SO₃ (100 mL, sat.). The resulting mixture was extracted with CH₂Cl₂ (1×1000 mL). The combined organic layers were washed with sat. Na₂CO₃ (aq.) (2×500 mL) and brine (1×500 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to afford ethyl (8R)-8-tert-butyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16),12,14-hexaene-5-carboxylate (75 g, 94.23%) as a brown solid.

Step 9: To a stirred solution of ethyl (8R)-8-tert-butyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (50.00 g, 126.437 mmol, 1.00 equiv.) in DCM (500.00 mL) was added BBr₃ (95.03 g, 379.326 mmol, 3.00 equiv.) dropwise at 0° C. under nitrogen atmosphere. The resulting mixture was stirred for 1 h at 40° C. under nitrogen atmosphere. The resulting mixture was poured into water (500 mL). The resulting mixture was extracted with CH₂Cl₂ (3×300 mL). The combined organic layers were washed with brine (1×500 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH₂Cl₂/MeOH (20:1) to afford ethyl (8R)-8-tert-butyl-15-hydroxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (25.1385 g, 52.13%) as a yellow solid. ¹H-NMR (DMSO-d6): δ 10.52 (s, 1H), 8.49 (s, 1H), 7.21 (dd, J=7.8, 1.2 Hz, 1H), 7.14 (t, J=7.8 Hz, 1H), 6.91 (dd, J=7.8, 1.2 Hz, 1H), 6.45 (s, 1H), 4.55 (d, J=6.4 Hz, 1H), 4.22 (q, J=7.0 Hz, 2H), 3.35 (dd, J=17.6, 6.6 Hz, 2H), 1.27 (t, J=7.1 Hz, 3H), 0.81 (s, 9H). ESI MS m/z=382.1 [M+H]⁺.

Step 10: A vial was charged with ethyl (8R)-8-tert-butyl-15-hydroxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2.7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (100 mg) and anhydrous DCM (20 mL) under a nitrogen atmosphere. Then, N-Phenyl-bis(trifluoromethanesulfonimide) (500 mg) was added and the reaction was stirred for 8 h at rt. The solvent was removed and the residue was purified on silica gel to provide ethyl (S)-6-(tert-butyl)-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (35 mg). ESI MS m/z=514.1 [M+H]⁺.

Step 11: An oven-dried vial was charged with ethyl (S)-6-(tert-butyl)-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (47 mg), 2-(furan-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (120 mg), Pd^(t)BuXPhos G3 (10 mg), and Cs₂CO₃ (194 mg). The vial was purged with nitrogen gas for 5 minutes, then DMF (3 mL) and water (0.8 mL) were added via syringe. The reaction mixture was heated under a nitrogen atmosphere for 80 minutes at 110° C. After cooling to room temperature, the reaction mixture was filtered and the product was purified by RPHPLC to provide ethyl (S)-6-(tert-butyl)-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as a white solid (10 mg). ESI MS m/z=432.1 [M+H]⁺.

Step 12: A vial was charged with ethyl (S)-6-(tert-butyl)-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (8 mg), MeOH (1 mL), THF (1 mL), 1 M aq. NaOH (0.5 mL). The reaction mixture was stirred for 4 h, then the organic solvents were removed under a stream of nitrogen. The pH of the mixture was adjusted to 3 by addition of 1 M aq. HCl. The product was extracted with DCM, then the DCM was removed under a stream of nitrogen. The residue was purified by RPHPLC to provide (S)-6-(tert-butyl)-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a white solid (1 mg). ESI MS m/z=404.1 [M+H]⁺.

Example 27: Synthesis of (S)-6-(tert-butyl)-11-(oxazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=405.1 [M+H]⁺.

Example 28: Synthesis of (S)-6-(tert-butyl)-11-(1-methyl-1H-pyrazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=418.1 [M+H]⁺.

Example 29: Synthesis of (S)-6-(tert-butyl)-11-(1-methyl-1H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=418.1 [M+H]⁺.

Example 30: Synthesis of (S)-6-(tert-butyl)-11-(1-methyl-1H-pyrazol-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=418.1 [M+H]⁺.

Example 31: Synthesis of (S)-6-(tert-butyl)-11-cyano-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=363.1 [M+H]⁺.

Example 32: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(1H-pyrazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=404.1 [M+H]⁺.

Example 33: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(1H-pyrazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylicacid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=404.1 [M+H]⁺.

Example 34: Synthesis of (S)-6-(tert-butyl)-11-(furan-2-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=404.1 [M+H]⁺.

Example 35: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(thiophen-3-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=420.1 [M+H]⁺.

Example 36: Synthesis of (S)-6-(tert-butyl)-11-(3,3-difluoropyrrolidin-1-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=443.1 [M+H]+.

Example 37: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(thiophen-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=420.1 [M+H]⁺.

Example 38: Synthesis of (S)-6-(tert-butyl)-11-(isothiazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=421.1 [M+H]⁺.

Example 39: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(thiazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=421.1 [M+H]⁺.

Example 40: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(pyrrolidin-1-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=407.1 [M+H]⁺.

Example 41: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(thiazol-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=421.1 [M+H]⁺.

Example 42: Synthesis of (S)-6-(tert-butyl)-2-oxo-11-(1H-pyrazol-1-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=404.1 [M+H]⁺.

Example 43: Synthesis of (S)-6-(tert-butyl)-11-cyclopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=378.1 [M+H]⁺.

Example 44: Synthesis of (S)-6-(tert-butyl)-11-methyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 26. ESI MS m/z=352.1 [M+H]⁺.

Example 45: Synthesis of (S)-6-(tert-butyl)-11-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: A vial was charged with ethyl (S)-6-(tert-butyl)-1-hydroxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (50 mg), Cs₂CO₃ (200 mg) and anhydrous DMF (20 mL) under a nitrogen atmosphere. Then, methyl iodide (0.1 mL) was added and the reaction was stirred for 8 h at 75° C. The reaction mixture was filtered and the filtrate was purified by RPHPLC to provide ethyl (S)-6-(tert-butyl)-11-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (12 mg). ESI MS m/z=396.1 [M+H]⁺.

Step 2: A vial was charged with ethyl (S)-6-(tert-butyl)-1-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (10 mg), MeOH (1 mL), THF (1 mL), 1 M aq. NaOH (0.5 mL). The reaction mixture was stirred for 4 h, then the organic solvents were removed under a stream of nitrogen. The pH of the mixture was adjusted to 3 by addition of 1 M aq. HCl. The product was extracted with DCM, then the DCM was removed under a stream of nitrogen. The residue was purified by RPHPLC to provide (S)-6-(tert-butyl)-11-methoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a white solid (5 mg). ESI MS m/z=368.1 [M+H]⁺.

Example 46: Synthesis of (S)-6-(tert-butyl)-11-(oxetan-3-yloxy)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 45. ESI MS m/z=410.1 [M+H]⁺.

Example 47: Synthesis of (S)-6-(tert-butyl)-11-cyclopropoxy-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 45. ESI MS m/z=410.1 [M+H]⁺.

Example 48: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: Into a 500-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed (4 S)-4-tert-butyl-10-methoxy-8-oxa-5-azatricyclo[7.4.0.0{circumflex over ( )}[2,7]]trideca-1(13),2(7),5,9,11-pentaene (20.00 g, 77.720 mmol, 1.00 equiv.), acetonitrile (200.00 mL), Zinciodide (24.81 g, 77.720 mmol, 1.00 equiv.), ethyl (2Z)-2-(ethoxymethylidene)-4,4-difluoro-3-[(trimethylsilyl)oxy]but-3-enoate (76.26 g, 155.441 mmol, 2.00 equiv., 60%). The resulting solution was stirred for 6 h at 50° C. in an oil bath. The reaction was then quenched by the addition of water/ice. The resulting solution was extracted with 3×500 mL of EA and the organic layers combined. The resulting mixture was washed with 2×500 ml of brine. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was applied onto a silica gel column with dichloromethane/methanol (50/1). This resulted in 15 g (46.68%) of ethyl (8S)-8-tert-butyl-3-fluoro-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11,13,15-hexaene-5-carboxylate as a black solid.

Step 2: Into a 250-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placedethyl(8 S)-8-tert-butyl-3-fluoro-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (15.00 g, 36.281 mmol, 1.00 equiv.), DCM (150.00 mL), and boron tribromide (27.27 g, 108.854 mmol, 3.00 equiv.) was added dropwise at 0° C. The resulting solution was stirred for 3 h at 0° C. in an ice/salt bath. The reaction was then quenched by the addition of water/ice. The pH value of the solution was adjusted to 8 with NaHCO₃(sat.). The resulting solution was extracted with 2×100 mL of dichloromethane. The residue was applied onto a flash gel column with ACN/Water (2/1). This resulted in 6.4 g (43.81%) of ethyl (8S)-8-tert-butyl-3-fluoro-15-hydroxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16),12,14-hexaene-5-carboxylate as a grey solid. ¹H-NMR (DMSO-d6): δ 10.35 (s, 1H), 8.56 (s, 1H), 7.28 (dd, J=6.0, 1.2 Hz, 1H), 7.18 (t, J=7.5 Hz, 1H), 6.93 (dd, J=9.0, 0.9 Hz, 1H), 4.66 (d, J=6.0 Hz, 1H), 4.25 (q, J=6.0 Hz, 2H), 3.42-3.39 (m, 2H), 1.29 (t, J=7.5 Hz, 3H), 0.79 (s, 9H). ESI MS m/z=400.1 [M+H]⁺.

Step 3: A vial was charged with ethyl (8S)-8-tert-butyl-3-fluoro-15-hydroxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0{circumflex over ( )}[2,7].0{circumflex over ( )}[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (100 mg) and anhydrous DCM (20 mL) under a nitrogen atmosphere. Then, N-Phenyl-bis(trifluoromethanesulfonimide) (500 mg) was added and the reaction was stirred for 8 h at rt. The solvent was removed and the residue was purified on silica gel to provide ethyl (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (54 mg). ESI MS m/z=532.1 [M+H]⁺.

Step 4: An oven-dried vial was charged with ethyl (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (54 mg), 2-(furan-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (122 mg), Pd^(t)BuXPhos G3 (10 mg), and Cs₂CO₃ (194 mg). The vial was purged with nitrogen gas for 5 minutes, then DMF (3 mL) and water (0.8 mL) were added via syringe. The reaction mixture was heated under a nitrogen atmosphere for 80 minutes at 110° C. After cooling to room temperature, the reaction mixture was filtered and the product was purified by RPHPLC to provide ethyl (S)-6-(tert-butyl)-1-fluoro-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as a white solid (18 mg). ESI MS m/z=450.1 [M+H]⁺.

Step 5: A vial was charged with ethyl (S)-6-(tert-butyl)-1-fluoro-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (18 mg), MeOH (1 mL), THF (1 mL), 1 M aq. NaOH (0.5 mL). The reaction mixture was stirred for 4 h, then the organic solvents were removed under a stream of nitrogen. The pH of the mixture was adjusted to 3 by addition of 1 M aq. HCl. The product was extracted with DCM, then the DCM was removed under a stream of nitrogen. The residue was purified by RPHPLC to provide (S)-6-(tert-butyl)-1-fluoro-11-(furan-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a white solid (6 mg). ESI MS m/z=422.1 [M+H]⁺.

Example 49: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(furan-2-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=422.1 [M+H]⁺.

Example 50: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(1H-pyrazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=422.1 [M+H]⁺.

Example 51: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(1H-pyrazol-5-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=422.1 [M+H]⁺.

Example 52: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(isothiazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=439.1 [M+H]⁺.

Example 53: Synthesis of (S)-6-(tert-butyl)-11-cyano-1-fluoro-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=381.1 [M+H]⁺.

Example 54: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(thiophen-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=438.1 [M+H]⁺.

Example 55: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(1-methyl-1H-pyrazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=436.1 [M+H]⁺.

Example 56: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(1-methyl-1H-pyrazol-3-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=436.1 [M+H]⁺.

Example 57: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(thiophen-3-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=438.1 [M+H]⁺.

Example 58: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(oxazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=423.1 [M+H]⁺.

Example 59: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-(1-methyl-1H-pyrazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=436.1 [M+H]⁺.

Example 60: Synthesis of (S)-6-(tert-butyl)-11-(3,3-difluoropyrrolidin-1-yl)-1-fluoro-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=461.1 [M+H]⁺.

Example 61: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(thiazol-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=439.1 [M+H]⁺.

Example 62: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(thiazol-4-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=439.1 [M+H]⁺.

Example 63: Synthesis of (S)-6-(tert-butyl)-1-fluoro-2-oxo-11-(1H-pyrazol-1-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=422.1 [M+H]⁺.

Example 64: Synthesis of (S)-6-(tert-butyl)-1-fluoro-11-methyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=370.1 [M+H]⁺.

Example 65: Synthesis of (S)-6-(tert-butyl)-11-cyclopropyl-1-fluoro-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 48. ESI MS m/z=396.1 [M+H]⁺.

Example 66: Synthesis of (S)-11-(furan-3-yl)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

Step 1: Into a 10 L 4-necked round-bottom flask were added ethyl crotonate (400.00 g, 3504.345 mmol, 1.00 equiv.), NBS (654.90 g, 3679.563 mmol, 1.05 equiv.), BPO (89.80 g, 350.435 mmol, 0.10 equiv.) and CCl₄ (5.00 L). The resulting mixture was stirred overnight at 80° C. under nitrogen atmosphere. The mixture was allowed to cool down to room temperature. The resulting mixture was filtered. The filtrate was washed with water (1×3 L) and brine (1×3 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (10:1) to afford ethyl (2E)-4-bromobut-2-enoate (400 g, 59.13%) as a colorless oil.

Step 2: Into a 10 L 4-necked round-bottom flask were added ethyl (2E)-4-bromobut-2-enoate (400.00 g, 2072.109 mmol, 1.00 equiv.), 2-bromo-6-methoxyphenol (420.71 g, 2072.109 mmol, 1.00 equiv.), K₂CO₃ (859.13 g, 6216.328 mmol, 3.00 equiv.) and DMF (4.00 L). The resulting mixture was stirred for 2 h at 60° C. The reaction was quenched by the addition of water (1 L). The resulting mixture was diluted with water (3 L). The resulting mixture was extracted with EtOAc (3×3 L). The combined organic layers were washed with brine (1×5 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (5:1) to afford ethyl (2E)-4-(2-bromo-6-methoxyphenoxy)but-2-enoate (480 g, 73.50%) as a yellow oil.

Step 3: Into a 10 L 4-necked round-bottom flask were added ethyl (2E)-4-(2-bromo-6-methoxyphenoxy)but-2-enoate (480.00 g, 1523.021 mmol, 1.00 equiv.), K₂CO₃ (631.47 g, 4569.064 mmol, 3.00 equiv.), Toluene (5.00 L) and Pd(PPh₃)₄ (52.80 g, 45.691 mmol, 0.03 equiv.). The resulting mixture was stirred for 4 h at 80° C. under nitrogen atmosphere. The reaction was quenched by the addition of water (2 L). The resulting mixture was extracted with EtOAc (3×3 L). The combined organic layers were washed with brine (1×4 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (10:1) to afford ethyl 2-(7-methoxy-1-benzofuran-3-yl)acetate (230 g, 64.47%) as a yellow oil.

Step 4: Into a 5 L 4-necked round-bottom flask were added ethyl 2-(7-methoxy-1-benzofuran-3-yl)acetate (230.00 g, 981.853 mmol, 1.00 equiv.), NaOH (117.81 g, 2945.464 mmol, 3.00 equiv.), THF (1.00 L) and H₂O (1.00 L). The resulting mixture was stirred for 1 h at room temperature. The resulting mixture was concentrated under reduced pressure. The mixture was acidified to pH 1-2 with HCl (6 N). The precipitated solids were collected by filtration and washed with water (1×1 L) to afford (7-methoxy-1-benzofuran-3-yl)acetic acid (150 g, 74.09%) as an off-white solid.

Step 5: To a stirred mixture of (7-methoxy-1-benzofuran-3-yl)acetic acid (150.00 g, 727.460 mmol, 1.00 equiv.) in DCM (500 mL) was added (COCl)₂ (277.00 g, 2182.379 mmol, 3.00 equiv.) dropwise at 0° C. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in THF (2.00 L). To the above mixture was added i-PrMgCl in THF (89.71 mL, 727.477 mmol, 1.00 equiv.) dropwise at 0° C. The resulting mixture was stirred for additional 30 min at room temperature. The reaction was quenched by the addition of sat. NH₄Cl (aq.) (500 mL). The resulting mixture was extracted with EtOAc (3×1 L). The combined organic layers were washed (1×1 L) with brine (1×1 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (20:1) to afford 1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-one (76 g, 44.98%) as a brown oil.

Step 6: To a stirred mixture of (S)-phenylethylamine (99.13 g, 817.982 mmol, 2.50 equiv.) and TEA (165.54 g, 1635.964 mmol, 5.00 equiv.) in DCE (1000.00 mL) was added TiCl₄ (62.06 g, 327.193 mmol, 1.00 equiv.) dropwise at 0° C. under nitrogen atmosphere. After 10 min, to the above mixture was added 1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-one (76.00 g, 327.193 mmol, 1.00 equiv.) dissolved in DCE (100 mL). The resulting mixture was stirred overnight at 70° C. The mixture was allowed to cool down to 0° C. To the above mixture was added NaBH₄ (24.76 g, 654.385 mmol, 2.00 equiv.) and MeOH (500.00 mL) in several portions. The resulting mixture was stirred for 3 h at room temperature. The reaction was quenched by the addition of water (500 mL). The mixture was acidified to pH 1-4 with 2N HCl. The resulting mixture was extracted with CH₂Cl₂ (3×500 mL). The combined organic layers were washed with Na₂CO₃ (aq.) (1×2 L) and brine (1×1 L), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE/EtOAc (30:1) to afford [(2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-yl][(1S)-1-phenylethyl]amine (90 g, 81.51%) as a brown oil.

Step 7: To a solution of [(2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-yl][(1S)-1-phenylethyl]amine (70.00 g, 207.430 mmol, 1.00 equiv.), D-Tartaric acid (31.13 g, 207.409 mmol, 1.00 equiv.) in MeOH (2.00 L) was added Pd(OH)₂/C (49.783 mmol, 0.24 equiv.) under nitrogen atmosphere in a 5 L 4-necked round-bottom flask. The mixture was hydrogenated at 30° C. for 3 h under hydrogen atmosphere using a hydrogen balloon, filtered through a Celite pad and concentrated under reduced pressure. The residue was dissolved in water (1 L). The mixture was basified to pH 8-9 with saturated Na₂CO₃ (aq.). The resulting mixture was extracted with CH₂Cl₂ (3×300 mL). The combined organic layers were washed with brine (1×500 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to afford (2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-amine (40 g, 82.65%) as a dark yellow oil.

Step 8: Into a 1 L 3-necked round-bottom flask were added (2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-amine (40.00 g, 171.445 mmol, 1.00 equiv.) and ethyl formate (400.00 mL). The resulting mixture was stirred overnight at 70° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure to afford N-[(2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-yl]formamide (40 g, 89.28%) as a brown yellow oil.

Step 9: Into a 1 L 3-necked round-bottom flask were added N-[(2S)-1-(7-methoxy-1-benzofuran-3-yl)-3-methylbutan-2-yl]formamide (40.00 g), methanesulfonic acid (100.00 mL) and ACN (300.00 mL). The resulting mixture was stirred for 5 h at 80° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (500 mL). The mixture was basified to pH 9-10 with saturated Na₂CO₃ (aq.). The resulting mixture was extracted with CH₂Cl₂ (3×300 mL). The combined organic layers were washed with brine (1×300 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to afford (4S)-4-isopropyl-10-methoxy-8-oxa-5-azatricyclo[7.4.0.0[2,7]]trideca-1(9),2(7),5,10,12-pentaene (32 g, 85.93%) as a brown oil.

Step 10: Into a 1 L 3-necked round-bottom flask were added (4S)-4-isopropyl-10-methoxy-8-oxa-5-azatricyclo[7.4.0.0[2,7]]trideca-1(9),2(7),5,10,12-pentaene (32.00 g, 131.522 mmol, 1.00 equiv.), ethyl (2E)-2-(ethoxymethylidene)-3-oxobutanoate (73.47 g, 394.565 mmol, 3.00 equiv.), TFA (74.98 g, 657.608 mmol, 5.00 equiv.) and EtOH (300.00 mL). The resulting mixture was stirred for 1 h at 80° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced to afford ethyl (8S)-8-isopropyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7]. 0[11,16]]heptadeca-1(10), 5,11(16), 12,14-pentaene-5-carboxylate (40 g, 79.32%) as a brown oil.

Step 11: To a stirred solution of ethyl (8S)-8-isopropyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0 [2,7].0[11,16]]heptadeca-1(10), 5,11(16),12,14-pentaene-5-carboxylate (40.00 g, 104.318 mmol, 1.00 equiv.) in DCM (200 mL) was added DDQ (28.40 g, 125.108 mmol, 1.20 equiv.). The resulting mixture was stirred for 1 h at room temperature. The reaction was quenched by the addition of sat. Na₂S₂O₃(aq.) (200 mL). The resulting mixture was extracted with CH₂Cl₂ (3×200 mL). The combined organic layers were washed with sat. Na₂CO₃ (aq.) (1×300 mL) and brine (1×300 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography with the following conditions: column, C18; mobile phase, ACN in water, 40% to 90% gradient in 25 min; detector, UV 254 nm. This resulted in ethyl (8S)-8-isopropyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7].0[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (35 g, 87.96%) as a brown solid.

Step 12: Into a 1 L 3-necked round-bottom flask were added ethyl (8S)-8-isopropyl-15-methoxy-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7]. 0[11,16]]heptadeca-1(10),2,5,11(16),12,14-hexaene-5-carboxylate (35.00 g, 91.760 mmol, 1.00 equiv.) and HBr in water (350.00 mL). The resulting mixture was stirred overnight at 100° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure to afford (8S)-15-hydroxy-8-isopropyl-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0 [2,7].0[11,16]]heptadeca-1(10),2,5,11(16),12,14-hexaene-5-carboxylic acid (22 g, 70.65%) as a brown solid.

Step 13: To a stirred solution of (8S)-15-hydroxy-8-isopropyl-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7].0[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylic acid (22.00 g, 64.830 mmol, 1.00 equiv.) in EtOH (250.00 mL) was added SOCl₂(23.14 g, 194.491 mmol, 3.00 equiv.) dropwise at 0° C. The resulting mixture was stirred overnight at 80° C. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with DCM (1000 mL). The resulting mixture was washed with 1×300 mL of saturated NaHCO₃(aq.). The combined organic layers were washed with brine (1×300 mL), dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse flash chromatography with the following conditions: column, C18; mobile phase, ACN in water, 10% to 90% gradient in 30 min; detector, UV 254 nm. This resulted in ethyl (8S)-15-hydroxy-8-isopropyl-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7].0[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (10.26 g, 43.08%) as a dark yellow solid. ¹H-NMR (DMSO-d6): δ 10.33 (s, 1H), 8.51 (s, 1H), 7.21 (dd, J=7.7, 1.3 Hz, 1H), 7.16 (t, J=7.7 Hz, 1H), 6.91 (dd, J=7.7, 1.3 Hz, 1H), 6.47 (s, 1H), 4.48 (dt, J=8.0, 3.8 Hz, 1H), 4.22 (qd, J=7.1, 2.0 Hz, 2H), 3.32 (d, J=3.9 Hz, 2H), 1.96 (q, J=7.0 Hz, 1H), 1.27 (t, J=7.1 Hz, 3H), 0.89 (d, J=6.6 Hz, 3H), 0.76 (d, J=6.8 Hz, 3H). ESI MS m/z=368.1 [M+H]⁺.

Step 14: A vial was charged with ethyl (8S)-15-hydroxy-8-isopropyl-4-oxo-17-oxa-7-azatetracyclo[8.7.0.0[2,7].0[11,16]]heptadeca-1(10),2,5,11(16), 12,14-hexaene-5-carboxylate (100 mg) and anhydrous DCM (20 mL) under a nitrogen atmosphere. Then, N-Phenyl-bis(trifluoromethanesulfonimide) (500 mg) was added and the reaction was stirred for 5 h at rt. The solvent was removed and the residue was purified on silica gel to provide ethyl (S)-6-isopropyl-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (34 mg). ESI MS m/z=500.1 [M+H]⁺.

Step 15: An oven-dried vial was charged with ethyl (S)-6-isopropyl-2-oxo-11-(((trifluoromethyl)sulfonyl)oxy)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (34 mg), 2-(furan-3-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (127 mg), Pd^(t)BuXPhos G3 (10 mg), and Cs₂CO₃ (194 mg). The vial was purged with nitrogen gas for 5 minutes, then DMF (3 mL) and water (0.8 mL) were added via syringe. The reaction mixture was heated under a nitrogen atmosphere for 80 minutes at 110° C. After cooling to room temperature, the reaction mixture was filtered and the product was purified by RPHPLC to provide ethyl (S)-11-(furan-3-yl)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate as a white solid (25 mg). ESI MS m/z=418.1 [M+H]⁺.

Step 5: A vial was charged with ethyl (S)-11-(furan-3-yl)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylate (25 mg), MeOH (1 mL), THF (1 mL), 1 M aq. NaOH (0.5 mL). The reaction mixture was stirred for 4 h, then the organic solvents were removed under a stream of nitrogen. The pH of the mixture was adjusted to 3 by addition of 1 M aq. HCl. The product was extracted with DCM, then the DCM was removed under a stream of nitrogen. The residue was purified by RPHPLC to provide (S)-1-(furan-3-yl)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid as a white solid (8 mg). ESI MS m/z=390.1 [M+H]⁺.

Example 67: Synthesis of (S)-11-(furan-2-yl)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=390.1 [M+H]⁺.

Example 68: Synthesis of (S)-10-(difluoromethoxy)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=390.1 [M+H]⁺.

Example 69: Synthesis of (S)-10-cyano-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=349.1 [M+H]⁺.

Example 70: Synthesis of (S)-6-isopropyl-11-(isothiazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=407.1 [M+H]⁺.

Example 71: Synthesis of (S)-6-isopropyl-11-(isothiazol-4-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=429.1 [M+H]⁺.

Example 72: Synthesis of (S)-6-isopropyl-2-oxo-10-(1H-pyrazol-1-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=390.1 [M+H]⁺.

Example 73: Synthesis of (S)-6-isopropyl-10-methyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=338.1 [M+H]⁺.

Example 74: Synthesis of (S)-10-cyclopropyl-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=364.1 [M+H]⁺.

Example 75: Synthesis of (S)-6-isopropyl-2-oxo-10-(thiophen-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=406.1 [M+H]⁺.

Example 76: Synthesis of (S)-6-isopropyl-2-oxo-10-(thiophen-3-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=406.1 [M+H]⁺.

Example 77: Synthesis of (S)-11-(difluoromethoxy)-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 4. ESI MS m/z=390.1 [M+H]⁺.

Example 78: Synthesis of (S)-11-cyano-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=349.1 [M+H]⁺.

Example 79: Synthesis of (S)-6-isopropyl-2-oxo-11-(thiophen-2-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=406.1 [M+H]⁺.

Example 80: Synthesis of (S)-6-isopropyl-2-oxo-11-(thiophen-3-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=406.1 [M+H]⁺.

Example 81: Synthesis of (S)-6-isopropyl-2-oxo-11-(1H-pyrazol-1-yl)-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=390.1 [M+H]⁺.

Example 82: Synthesis of (S)-6-isopropyl-11-methyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=338.1 [M+H]⁺.

Example 83: Synthesis of (S)-11-cyclopropyl-6-isopropyl-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=364.1 [M+H]⁺.

Example 84: Synthesis of (S)-6-isopropyl-11-(oxazol-5-yl)-2-oxo-6,7-dihydro-2H-benzofuro[2,3-a]quinolizine-3-carboxylic acid

The title compound was prepared in a manner analogous to the methods used for Example 65. ESI MS m/z=391.1 [M+H]⁺.

Biological Activity

Methods:

2.2.15 cells are passaged upon attaining confluency in DMEM/F12 media in the presence of 10% FBS, Penn/Strep, and 250 ug/mL G418. Novel compounds are 5 fold serially diluted in DMSO and added to 96 well plates containing 35,000 cells/well at a 1:200 dilution so that the final concentration of DMSO is 0.5%. On day 5, post treatment cell lysates and supernatants are harvested for analysis.

Cells are lysed using Agilent Sidestep Lysis buffer, diluted 1:100 and quantified via quantitative real time PCR. Commercially available ELISA kits are used to quantitate the viral proteins HBsAg (Alpco) or HBeAg (US Biological) by following the manufacturer's recommended protocol after diluting samples to match the linear range of their respective assays. Irrespective of readout, compound concentrations that reduce viral product accumulation in the cell lysates or supernatants by 50% relative to no drug controls (EC₅₀) are reported; EC₅₀ ranges are as follows: A<0.1 μM; B 0.1-1 μM; C>1 μM.

Additionally, compound induced cellular toxicity is evaluated by exposing HepG2 cells seeded at 5,000 cells/well to serially diluted compound with a final DMSO concentration of 0.5% for three days. At day 3, post seeding cells are treated with ATPlite 1 Step according to the manufacturer's instructions. Compound concentrations that reduce total ATP levels in wells by 50% relative to no drug controls (CC₅₀) are reported; CC₅₀ ranges are as follows: A>25 μM; B 10-25 μM; C<10 μM.

TABLE 2 Summary of Activities Example 2.2.15 cells HepG2 cells Number EC₅₀ (μM) CC₅₀ (μM) 1 B A 2 C A 3 A A 4 A A 5 A — 6 A B 7 A A 8 A A 9 A — 10 A A 11 A A 12 A A 13 A A 14 A A 15 A — 16 A — 17 A A 18 A — 19 A — 20 A — 21 A B 22 A — 23 A B 24 A A 25 A A 26 A B 27 A B 28 A A 29 A B 30 A B 31 A A 32 A A 33 A A 34 A B 35 A A 36 A A 37 A A 38 A B 39 A B 40 A — 41 A B 42 A A 43 A B 44 B — 45 A A 46 A A 47 A B 48 A A 49 A A 50 A A 51 A A 52 A A 53 A A 54 A A 55 A A 56 A — 57 A A 58 A A 59 A A 60 A — 61 A A 62 A B 63 A A 64 A A 65 A A 66 A B 67 A B 68 A C 69 A A 70 A A 71 A C 72 A C 73 A A 74 A B 75 A — 76 A A 77 A A 78 A A 79 A C 80 A C 81 A A 82 A A 83 A C 84 A A

While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims. 

What is claimed:
 1. A compound represented by Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: Z₁ is O, NR₁, or S; Z₂ is N or CR₂; Z₃ is N or CR₃; Z₄ is N or CR₄; Z₅ is N or CR₅; R₁ is hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; R₂, R₃, R₄, and R₅ are each independently selected from: 1) hydrogen; 2) halogen; 3) —NO₂; 4) Cyano; 5) Optionally substituted —C₁-C₈ alkyl; 6) Optionally substituted —C₂-C₈ alkenyl; 7) Optionally substituted —C₂-C₈ alkynyl; 8) Optionally substituted —C₃-C₈ cycloalkyl; 9) Optionally substituted 3- to 8-membered heterocycloalkyl; 10) Optionally substituted aryl; 11) Optionally substituted arylalkyl; 12) Optionally substituted heteroaryl; 13) Optionally substituted heteroarylalkyl; 14) —SR₁₁; 15) —S(O)₂R₁₁; 16) —S(O)₂N(R₁₁)(R₁₂); 17) —C(O)R₁₁; 18) —C(O)OR₁₁; 19) —C(O)N(R₁₁)(R₁₂); 20) —C(O)N(R₁₁)S(O)₂(R₁₂); 21) —N(R₁₁)(R₁₂); 22) —N(R₁₃)C(O) N(R₁₁)(R₁₂); 23) —N(R₁₁)C(O)(R₁₂); 24) —N(R₁₁)C(O)₂(R₁₂); 25) —N(R₁₃) S(O)₂N(R₁₁)(R₁₂); 26) —N(R₁₁)S(O)₂(R₁₂); 27) —OR₁₁; 28) —OC(O)R₁₁; 29) —OC(O)OR₁₁; and 30) —OC(O)N(R₁₁)(R₁₂); wherein R₁₁, R₁₂, and R₁₃ are each independently selected from hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; alternatively, R₁₁ and R₁₂ are taken together with the nitrogen atom to which they attached to form an optionally substituted 3-8 membered heterocyclic containing 0, 1, 2, or 3 double bonds; alternatively, Z₂ is CR₂, Z₃ is CR₃, and R₂ and R₃ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Z₃ is CR₃, Z₄ is CR₄, and R₃ and R₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Z₄ is CR₄, Z₅ is CR₅, and R₄ and R₅ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; Q₁, Q₂, and Q₃ are each independently selected from hydrogen, halo, NR₁₁, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈ cycloalkyl; optionally substituted 3- to 8-membered heterocycloalkyl; optionally substituted aryl; and optionally substituted heteroaryl; Q₄ is selected from halo, NR₁₁, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈ cycloalkyl; optionally substituted 3- to 8-membered heterocycloalkyl optionally substituted aryl; and optionally substituted heteroaryl; alternatively, Q₁ is taken together with Q₂ or Q₃ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Q₂ is taken together with Q₄ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Q₃ and Q₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; Y₁ is hydrogen, halo, or optionally substituted C₁-C₆ alkyl; Y₂ is O, NR₁₁, N(OR₁₁), or N(NR₁₁); Y₃ is —COOR₁₁, —C(O)NHSO₂R₁₁, —C(O)NHSO₂NR₁₁R₁₂, 5-tetrazolyl, or 1,2,4-oxadiazol-3-yl-5(4H)-one; and Y₄ is hydrogen or optionally substituted methyl; alternatively, Y₂ and Y₃ are taken together with the carbon atoms to which they are attached to form an optionally substituted 5-12 membered heterocyclic ring containing 1, 2, or 3 double bonds.
 2. The compound of claim 1, wherein at least one of Z₂, Z₃, Z₄ and Z₅ is not nitrogen and at least one of R₂, R₃, R₄, and R₅ is selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic, wherein R₁₁ and R₁₂ are as defined in claim
 1. 3. The compound of claim 1, wherein at least one of Z₂, Z₃, Z₄ and Z₅ is not nitrogen and at least one of R₂, R₃, R₄, and R₅ is selected from hydrogen, halogen, —CN, —CH₃, —CF₃, —CHF₂, —C(O)CH₃, —OCH₃, —OCF₃, —OCHF₂, —OH, —OR₁₁, —NH₂, and —NHR₁₂, wherein R₁₁ and R₁₂ are each independently selected from one of the following by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic.
 4. The compound of claim 1, wherein at least one of Z₂, Z₃, Z₄ and Z₅ is not nitrogen and at least one of R₂, R₃, R₄, and R₅ is selected from

wherein each R₂₁ is independently selected from —CH₃, —CHF₂, —CF₃, -isopropyl, and -t-butyl; or at least one of R₂, R₃, R₄, and R₅ is selected from one of the following groups by removal of a hydrogen atom:

wherein each of these groups is optionally substituted with one to four groups selected from halo, CN, —OR₁₁, —NR₁₁R₁₂, optionally substituted C₁-C₆ alkyl, and optionally substituted 3- to 8-membered heterocyclic, wherein R₁₁ and R₁₂ are as defined in claim
 1. 5. The compound of claim 1, represented by Formula (II), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, Z₅, Q₁, Q₂, Q₃, and Q₄ are as defined in claim
 1. 6. The compound of claim 1, represented by Formula (IV-1) or Formula (IV-2), or a pharmaceutically acceptable salt thereof:

wherein Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ are as defined in claim
 1. 7. The compound of claim 1, represented by Formula (VI), or a pharmaceutically acceptable salt thereof:

wherein one V is —O—, —C(O)—, —S—, —S(O)₂—, —NR₂₂— or —C(R₂₂)₂—, and the other Vs are independently —O—, —NR₂₂— or —C(R₂₂)₂—; each R₂₂ is independently hydrogen, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₂-C₆ alkenyl, optionally substituted-C₂-C₆ alkynyl, optionally substituted C₁-C₆ alkoxy; optionally substituted —C₃-C₇ cycloalkyl, optionally substituted 3- to 7-membered heterocyclic, optionally substituted aryl or optionally substituted heteroaryl; n is 0, 1, 2 or 3; and Y₁, Y₃, Y₄, Z₁, Z₂, Z₃, Z₄, and Z₅ are as defined in claim
 1. 8. The compound of claim 1, represented by one of Formulae (VIII-1)˜(VIII-6) and Formulae (IX-1)˜(IX-6), or a pharmaceutically acceptable salt thereof:

wherein one V is —O—, —C(O)—, —S—, —S(O)₂—, —NR₂₂— or —C(R₂₂)₂—, and the other Vs are independently —O—, —NR₂₂— or —C(R₂₂)₂—; each R₂₂ is independently hydrogen, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₂-C₆ alkenyl, optionally substituted —C₂-C₆ alkynyl, optionally substituted C₁-C₆ alkoxy; optionally substituted —C₃-C₇ cycloalkyl, optionally substituted 3- to 7-membered heterocyclic, optionally substituted aryl or optionally substituted heteroaryl; and Y₁, Y₃, Y₄, R₁, R₂, R₃, Z₂, Z₃, Z₄, and Z₅ are as defined in claim
 1. 9. The compound of claim 1, represented by one of Formulae (XIII-1) or (XIII-2), or a pharmaceutically acceptable salt thereof:

wherein R₂, R₃, and Q₄ are as are as defined in claim
 1. 10. A compound selected from the compounds set forth below or a pharmaceutically acceptable salt thereof: Compound Structure 1

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11. A pharmaceutical composition, comprising a compound represented by Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: Z₁ is O, NR₁, or S; Z₂ is N or CR₂; Z₃ is N or CR₃; Z₄ is N or CR₄; Z₅ is N or CR₅; R₁ is hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; R₂, R₃, R₄, and R₅ are each independently selected from: 1) hydrogen; 2) halogen; 3) —NO₂; 4) Cyano; 5) Optionally substituted —C₁-C₈ alkyl; 6) Optionally substituted —C₂-C₈ alkenyl; 7) Optionally substituted —C₂-C₈ alkynyl; 8) Optionally substituted —C₃-C₈ cycloalkyl; 9) Optionally substituted 3- to 8-membered heterocycloalkyl; 10) Optionally substituted aryl; 11) Optionally substituted arylalkyl; 12) Optionally substituted heteroaryl; 13) Optionally substituted heteroarylalkyl; 14) —SR₁₁; 15) —S(O)₂R₁₁; 16) —S(O)₂N(R₁₁)(R₁₂); 17) —C(O)R₁₁; 18) —C(O)OR₁₁; 19) —C(O)N(R₁₁)(R₁₂); 20) —C(O)N(R₁₁)S(O)₂(R₁₂); 21) —N(R₁₁)(R₁₂); 22) —N(R₁₃)C(O) N(R₁₁)(R₁₂); 23) —N(R₁₁)C(O)(R₁₂); 24) —N(R₁₁)C(O)₂(R₁₂); 25) —N(R₁₃) S(O)₂N(R₁₁)(R₁₂); 26) —N(R₁₁)S(O)₂(R₁₂); 27) —OR₁₁; 28) —OC(O)R₁₁; 29) —OC(O)OR₁₁; and 30) —OC(O)N(R₁₁)(R₁₂); wherein R₁₁, R₁₂, and R₁₃ are each independently selected from hydrogen, optionally substituted —C₁-C₈ alkyl, optionally substituted —C₂-C₈ alkenyl, optionally substituted —C₃-C₈ cycloalkyl, optionally substituted 3- to 8-membered heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl: alternatively, R₁₁ and R₁₂ are taken together with the nitrogen atom to which they attached to form an optionally substituted 3-8 membered heterocyclic containing 0, 1, 2, or 3 double bonds; alternatively, Z₂ is CR₂, Z₃ is CR₃, and R₂ and R₃ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Z₃ is CR₃, Z₄ is CR₄, and R₃ and R₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Z₄ is CR₄, Z₅ is CR₅, and R₄ and R₅ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; Q₁, Q₂, Q₃ and Q₄ are each independently selected from hydrogen, halo, NR₁₁, optionally substituted —C₁-C₆ alkyl, optionally substituted —C₁-C₆ alkoxy, optionally substituted —C₃-C₈ cycloalkyl; optionally substituted 3- to 8-membered heterocycloalkyl: optionally substituted aryl; and optionally substituted heteroaryl; alternatively, Q₁ is taken together with Q₂ or Q₃ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Q₂ is taken together with Q₄ and the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; alternatively, Q₃ and Q₄ are taken together with the carbon atoms to which they are attached to form an optionally substituted 3-8 membered heterocyclic or carbocyclic ring containing 0, 1, 2, or 3 double bonds; Y₁ is hydrogen, halo, or optionally substituted C₁-C₆ alkyl; Y₂ is O, NR₁₁, N(OR₁₁), or N(NR₁₁); Y₃ is —COOR₁₁, —C(O)NHSO₂R₁₁, —C(O)NHSO₂NR₁₁R₁₂, 5-tetrazolyl, or 1,2,4-oxadiazol-3-yl-5(4H)-one; and Y₄ is hydrogen or optionally substituted methyl; alternatively, Y₂ and Y₃ are taken together with the carbon atoms to which they are attached to form an optionally substituted 5-12 membered heterocyclic ring containing 1, 2, or 3 double bonds; and a pharmaceutically acceptable carrier or excipient.
 12. A method of treating or preventing an HBV infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition according to claim
 11. 13. The method of claim 12, further comprising administering to the subject an additional therapeutic agent selected from the group consisting of a HBV polymerase inhibitor, interferon, viral entry inhibitor, viral maturation inhibitor, literature-described capsid assembly modulator, reverse transcriptase inhibitor, TLR-agonist, inducer of cellular viral RNA sensor, therapeutic vaccine, and agents of distinct or unknown mechanism, and a combination thereof.
 14. The method of claim 13, wherein the pharmaceutical composition comprises the additional therapeutic agent.
 15. The method of claim 13, wherein the compound and the additional therapeutic agent are co-administered.
 16. The method of claim 13, wherein the additional therapeutic agent is administered at a lower dose or frequency compared to the dose or frequency of the additional therapeutic agent that is required to treat an HBV infection when administered alone.
 17. The method of claim 13, wherein the subject is refractory to at least one compound selected from the group consisting of a HBV polymerase inhibitor, interferon, viral entry inhibitor, viral maturation inhibitor, distinct capsid assembly modulator, inducer of cellular viral RNA sensor, therapeutic vaccine, antiviral compounds of distinct or unknown mechanism, and combination thereof. 